Fused tricyclic dual inhibitors of cdk 4/6 and flt3

ABSTRACT

Compounds of Formula I are useful inhibitors of CDK 4, CDK6, and FLT3. Such compounds are useful in treating cancer and various other disease conditions. Compounds of Formula I have the following structure: 
     
       
         
         
             
             
         
       
         
         
           
             where R 1  is a group of Formula IA, Formula IB, Formula IC, or Formula ID 
           
         
       
    
     
       
         
         
             
             
         
       
     
     and the definitions of the other variables are provided herein.

CROSS REFERENCES TO RELATED APPLICATIONS

This application claims the benefit of U.S. Provisional Application No.61/466,841, filed on Mar. 23, 2011, which is hereby incorporated byreference in its entirety and for all purposes as if fully set forthherein.

FIELD OF THE INVENTION

The present invention relates to compounds capable of inhibiting thekinase activity of cyclin dependent kinases such as CDK4 and CDK6, andcompositions that include compounds that inhibit cyclin dependentkinases. The present invention also relates to compounds capable ofinhibiting the kinase activity of FLT3, and compositions that includecompounds that inhibit FLT3. The compounds and compositions may be usedto treat various diseases or conditions modulated by these kinases suchas cancer and are especially useful in treating patients with cancers.

BACKGROUND OF THE INVENTION

Acute myeloid leukemia (AML) represents a significant unmet medicalneed. It is a hematological malignancy characterized by a block indifferentiation and aberrant proliferation of the myeloid lineage ofhematopoietic progenitor cells. There are approximately 13,000 new casesand 9,000 deaths per year in the United States. The survival rate is25-70% in patients younger than 60 years and 5-15% in older patients,with worse outcomes in patients with poor risk cytogenetics. Currentstandard of care treatment is daunorubicin and cytarabine chemotherapywith induction and consolidation phases. Bone marrow stem celltransplant is also used for treating AML in younger patients.

Cyclin-dependent kinases (CDKs) are a family of serine/threonine proteinkinases playing important cellular functions. The cyclins are theregulatory subunits that activate the catalytic CDKs. CDK1/Cyclin B1,CDK2/Cyclin A, CDK2/Cyclin E, CDK4/Cyclin D, CDK6/Cyclin D are criticalregulators of cell cycle progression. CDKs also regulate transcription,DNA repair, differentiation, senescence and apoptosis (Morgan, D. O.,Annu. Rev. Cell. Dev. Biol., 13:261-291 (1997)).

Small molecule inhibitors of CDKs have been developed to treat cancer(de Carcer, G. et al., Curr. Med. Chem., 14:969-85 (2007)). A largeamount of genetic evidence supports that CDKs, their substrates orregulators have been shown to be associated with many human cancers(Malumbres, M. et al, Nature Rev. Cancer, 1:222-231 (2001)). Endogenousprotein inhibitors of CDKs including p16, p21 and p27 inhibit CDKactivity and their overexpression results in cell cycle arrest andinhibition of tumor growth in preclinical models (Kamb, A., Curr. Top.Microbiolo. Immunol., 227:139-148 (1998)).

Small molecule inhibitors of CDKs may also be used to treat variety ofother diseases that result from aberrant cell proliferation, includingcardiovascular disorders, renal diseases, certain infectious diseasesand autoimmune diseases. Cell proliferation pathways including genesinvolved in the cell cycle G1 and S phase checkpoint (p53, pRb, p15,p16, and Cyclins A, D, E, CDK 2 and CDK4) have been associated withplaque progression, stenosis and restenosis after angioplasty.Over-expression of the CDK inhibitor protein p21 has been shown toinhibit vascular smooth muscle proliferation and intimal hyperplasiafollowing angioplasty (Chang, M. W. et al., J. Clin. Invest., 96:2260(1995); Yang, Z-Y. et al., Proc. Natl. Acad. Sci. (USA) 93:9905 (1996)).A small molecule CDK2 inhibitor CVT-313 (Ki=95 nM) was shown to causesignificant inhibition of neointima formation in animal models (Brooks,E. E. et al., J. Biol. Chem., 272:29207-29211 (1997)). Disregulation ofcell cycle has been associated with polycystic kidney diseases, whichare characterized by the growth of fluid-filled cysts in renal tubules.Treatment with small molecule inhibitors of CDKs yielded effectivearrest of cystic disease in mouse models (Bukanov, N. O., et al.,Nature, 4444:949-952 (2006)). Infection by a variety of infectiousagents, including fungi, protozoan parasites such as Plasmodiumfalciparum, and DNA and RNA viruses may be treated with CDK inhibitors.CDKs have been shown to be required for replication of herpes simplexvirus (HSV) (Schang, L. M. et al., J. Virol., 72:5626 (1998)). Synovialtissue hyperplasia plays important roles in the development ofrheumatoid arthritis; inhibition of synovial tissue proliferation maysuppress inflammation and prevent joint destruction. It has been shownthat over-expression of CDK inhibitor protein p16 inhibited synovialfibroblast growth (Taniguchi, K. et al., Nat. Med., 5:760-767 (1999))and joint swelling was substantially inhibited in animal arthritismodels.

Selective inhibitors of some CDKs may also be used to protect normaluntransformed cells by inhibiting specific phases of cell cycleprogression (Chen, et al., J. Natl. Cancer Institute, 92:1999-2008(2000)). Pre-treatment with a selective CDK inhibitor prior to the useof a cytotoxic agent that inhibits a different phase of the cell cyclemay reduce the side effects associated with the cytotoxic chemotherapyand possibly increase the therapeutic widow. It has been shown thatinduction of cellular protein inhibitors of CDKs (p16, p27 and p21)conferred strong resistance to paclitaxel- or cisplatin-mediatedcytotoxicity on the inhibitor-responsive cells but not on theinhibitor-unresponsive cells (Schmidt, M, Oncogene, 2001 20:6164-71).

CDK4 and CDK6 are two functionally indistinguishable cyclin D dependentkinases. They are widely expressed with high levels of expressionobserved in cells of hematopoeitic lineage (CDK4/6 will be usedthroughout this document to reference both CDK4 and CDK6). CDK4/6promotes G1-S transition of the cell cycle by phosphorylating theretinoblastoma protein (Rb). CDK4 and CDK6 single knockout mice areviable and double knockout mice die around birth with defectivehematopoiesis (Satyanarayana, A. et al., Oncogene, 28:2925-39 (2009);Malumbres, M. et al., Cell, 118:493-504 (2004)). Strong evidencesupports a significant involvement of the cyclin D-CDK4-p16^(INK4A)-Rbpathway in cancer development (Malumbres, M. et al., Nature Rev. Cancer,1:222-31 (2001)). Rb negatively regulates the cell cycle at G1 bysequestering E2F proteins that are required for initiation of S phase.p16^(INK4A) is a key member of the INK4 family of CDK4/6 cellularinhibitors. The genes for Rb and p16^(INK4A) are tumor suppressors thatare often deleted or silenced in cancer cells. Additionally CDK4, CDK6and cyclin D are reported to be amplified in hematologic malignanciesand solid tumors. The importance of this pathway in oncogenesis isfurther supported by the finding that depletion or inactivation of CDK4inhibits tumor growth in mouse tumor models (Yu, Q. et al., Cancer Cell,9:23-32 (2006); Puyol, M. Cancer Cell, 18:63-73 (2010)). Rb andp16^(INK4A) are rarely deleted in AML. However, the p15^(INK4B) gene,another member of the INK4 family, has been reported to be downregulated by hypermethylation in up to 60% of AML (Naofumi, M. et al.,Leukemia Res., 29:557-64 (2005); Drexler, H. G. Leukemia, 12:845-59(1998); Herman, J. G. et al., Cancer Res., 57:837-41 (1997)), suggestinga possible critical role for CDK4/6 in AML cells.

FLT3 (Fms-like tyrosine kinase 3, FLK2) is a class III receptor tyrosinekinase. It is activated by the FLT3 ligand (FL) and signals through thePI3K, RAS, and JAK/STAT pathways (Scholl C. et al., Semin. Oncol.,35:336-45 (2008); Meshinchi S. et al., Clin. Cancer Res., 15:4263-9(2009)). FLT3 plays a role in early hematopoiesis and FLT3 deficientmice have reduced numbers of progenitors of multiple lymphoid lineages(Mackarehtschian K, et al., Immunity, 3:147-61 (1995). Activatingmutations in FLT3 are found in approximately 30% of AML patients,representing the most frequent genetic alteration in the disease. About75% of the activating mutations are internal tandem duplications (ITD)and 25% are point mutations in the activation loop of the kinase domain.The most frequently identified activating point mutation is D835Y(Yamamoto et al., Blood, 97(8): 2434-2439 (2001)). However, mutationshave also been found at N841I (Jiang, J. et al., Blood, 104(6):1855-1858 (2004)) and Y842C (Kindler et al., Blood, 105(1): 335-340(2005)). Additional point mutations have been identified in thejuxtamembrane domain and kinase domain, although these have been shownto result in lower transforming potential (Reindel et al., Blood 107(9):3700-3707 (2006)).

Murine bone marrow transplanted with a retrovirus expressing theFLT3-ITD has been shown to result in the production of a lethalmyeloproliferative disease in mice (Kelly et al., Blood 99: 310-318(2002)) characterized by leukocytosis consisting of mature neutrophils.This disease did not show a block in differentiation as seen in humanAML suggesting that FLT3 mutations confer a proliferative or survivaladvantage to the cells. Additional oncogene mutation producing a blockin differentiation such as AML1/ETO is hypothesized to be required toproduce disease that is more similar to human AML.

A number of FLT3 inhibitors have been tested in clinical trials.Although they have shown initial clinical responses in AML, theresponses observed were transient and resistance can develop rapidly(Weisberg, E. et al., Oncogene, 29:5120-34 (2010)). The major resistancemechanism appears to be through the acquisition of secondary mutationsin FLT3, which may interfere with the binding of FLT3 inhibitors to theFLT3 receptor (Weisberg, E. et al., Oncogene, 29:5120-34 (2010); Chu, S.H. et al., Drug Resist. Update, 12:8-16 (2009)). One such resistancemutation (N676K) was identified in a patient at the time of clinicalrelapse while on multi-kinase FLT3 inhibitor midostaurin (PKC412)monotherapy (Heidel, F. et al., Blood, 107:293-300 (2006)). Combinationsof FLT3 inhibitors with chemotherapy are being tested in clinical trialsdespite the recognition that chemotherapy is poorly tolerated.Additional possible mechanisms for lack of durable responses includeinadequate target coverage (Pratz, K. W., et al., Blood, 139:3938-46(2009)) and protection of AML cells in the bone marrow where stromalgrowth factors may provide proliferative signals in addition to FLT3activation (Tam, W. F. et al., Best Pract. Res. Clin. Haematol.,21:13-20 (2008)) Inhibitors with combined FLT3 and CDK4/6 inhibitoryactivities are novel and may prove beneficial in treating variouscancers including, but not limited to, AML.

Fused tricyclic pyridine, pyrimidine, and triazine compounds useful fortreating diseases mediated by CDK4 are disclosed in WO 2009/085185,published on Jul. 9, 2009, which is hereby incorporated by reference inits entirety and for all purposes as if fully set forth herein. Variousgem-disubstituted and spirocyclic compounds useful for treating diseasesmediated by CDK4 are disclosed in WO 2009/0126584, published on Oct. 15,2009, which is hereby incorporated by reference in its entirety and forall purposes as if fully set forth herein.

A continued need exists for new compounds that can be used to modulateCDK4, CDK6, and/or FLT3 and can be used to treat various diseaseconditions associated with these kinases. The compounds of the presentinvention provide significant improvements in inhibition in one or moreof these kinases and have properties making them excellent therapeuticcandidates.

SUMMARY OF THE INVENTION

In one aspect, the invention provides a compound of Formula I:

or a pharmaceutically acceptable salt thereof, a hydrate thereof, or amixture thereof, wherein:

R¹ is a group of Formula IA, Formula IB, Formula IC, or Formula ID

wherein the

symbol indicates the point of attachment of the group of Formula IA, IB,IC, or ID to the rest of the molecule;

R² is a C₅-C₇ cycloalkyl group, is a 5 to 7-membered heterocyclyl groupthat includes 1, 2, or 3 heteroatoms selected from N, O, and S, or is aC₇-C₁₀ bicyclic group; wherein the C₅-C₇ cycloalkyl group, the 5 to 7membered heterocyclyl group, or the C₇-C₁₀ bicyclic group isunsubstituted or is substituted with 1-3 substituents independentlyselected from unsubstituted —(C₁-C₆ alkyl), —OH, halo, —O—(C₁-C₆ alkyl),—CO₂H, —C(═O)—O—(C₁-C₆ alkyl), —C(═O)—NR′R″, —NR′R″, or a substituted—(C₁-C₄ alkyl), wherein the substituted —(C₁-C₄ alkyl) is substitutedwith 1-3 substituents independently selected from halo, —OH, —OCH₃,—S(═O)₂—CH₃, or —C(═O)—CH₃;

R^(3a) is selected from —H, —F, or —Cl, —(C₁-C₃ alkyl), or —O—(C₁-C₃alkyl);

R^(3b) is —H, halo, —OH, —O—(C₁-C₆ alkyl), unsubstituted —(C₁-C₆ alkyl),—NR′R″, —C(═O)—(C₁-C₆ alkyl), —C(═O)—O—(C₁-C₆ alkyl), —C(═O)—NR′R″, or asubstituted —(C₁-C₆ alkyl), wherein the substituted —(C₁-C₆ alkyl) issubstituted with 1-3 substituents independently selected from halo, —OH,—OCH₃, —CN, or —NO₂;

R^(3c) is —H, —(C₁-C₃ alkyl), or halo;

R⁴ is —H;

R⁵ is —H;

R⁶ is selected from —H, —(C₁-C₆ alkyl), —C(═O)—(C₁-C₆ alkyl),—C(═O)—O—(C₁-C₆ alkyl), —C(═O)—C(═O)—OH, —C(═O)—NR′R″, or —S(═O)—NR′R″,wherein the alkyl group of the —(C₁-C₆ alkyl), —C(═O)—(C₁-C₆ alkyl), and—C(═O)—O—(C₁-C₆ alkyl) groups is unsubstituted or is substituted with1-3 substituents independently selected from —OH, F, —S(═O)₂—(C₁-C₆alkyl), —O—(C₁-C₆ alkyl), —NR′R″, or —CN;

R^(7a) is —H, —CH₃, or halo;

R^(7b) is —H, —(C₁-C₆ alkyl), or halo; or R^(7b) is absent if R¹ is agroup of Formula IB or Formula ID;

R^(7c) is —H, unsubstituted —(C₁-C₆ alkyl), halo, —O—(C₁-C₆ alkyl),—NO₂, —CN, —NR′R″, —CO₂H, —C(═O)—O—(C₁-C₆ alkyl), —C(═O)—NR′R″, or asubstituted —(C₁-C₆ alkyl), wherein the substituted —(C₁-C₆ alkyl) issubstituted with 1-3 substituents independently selected from —OH, halo,—O—(C₁-C₆ alkyl), —CN, —NR′R″, or —S(═O)₂—CH₃; or R^(7c) is absent if R¹is a group of Formula IA or Formula IC;

R^(8a) is —H, unsubstituted —(C₁-C₆ alkyl), or a substituted —(C₁-C₆alkyl), wherein the substituted —(C₁-C₆ alkyl) is substituted with 1-3substituents independently selected from —OH, halo, or —O—(C₁-C₆ alkyl);

R^(8b) is —H, unsubstituted —(C₁-C₆ alkyl), or a substituted —(C₁-C₆alkyl), wherein the substituted —(C₁-C₆ alkyl) is substituted with 1-3substituents independently selected from —OH, halo, or —O—(C₁-C₆ alkyl);or R^(8a) and R^(8b), when taken together, can represent ═O;

R^(8c) is selected from —H, —OH, unsubstituted —(C₁-C₆ alkyl), or asubstituted —(C₁-C₆ alkyl), wherein the substituted —(C₁-C₆ alkyl) issubstituted with 1-3 substituents independently selected from —OH, halo,or —O—(C₁-C₆ alkyl);

R^(8d) is —H, unsubstituted —(C₁-C₆ alkyl), or a substituted —(C₁-C₆alkyl), wherein the substituted —(C₁-C₆ alkyl) is substituted with 1-3substituents independently selected from —OH, halo, or —O—(C₁-C₆ alkyl);

R^(8e) is —H, unsubstituted —(C₁-C₆ alkyl), or a substituted —(C₁-C₆alkyl), wherein the substituted —(C₁-C₆ alkyl) is substituted with 1-3substituents independently selected from —OH, halo, or —O—(C₁-C₆ alkyl);

R^(8f) is —H, unsubstituted —(C₁-C₆ alkyl), or a substituted —(C₁-C₆alkyl), wherein the substituted —(C₁-C₆ alkyl) is substituted with 1-3substituents independently selected from —OH, halo, or —O—(C₁-C₆ alkyl);or R^(8e) and R^(8f), when taken together, can represent ═O; and

R′ and R″ are independently selected from —H, unsubstituted —(C₁-C₄alkyl), or —(C₁-C₄ alkyl) substituted with 1 to 3 substituentsindependently selected from —OH or —F.

In some embodiments of the compound of Formula I or the pharmaceuticallyacceptable salt thereof, the hydrate thereof, or the mixture thereof, R²is a C₅-C₇ cycloalkyl group that is unsubstituted or is substituted with1-3 substituents independently selected from unsubstituted —(C₁-C₆alkyl), —OH, halo, —O—(C₁-C₆ alkyl), —CO₂H, —C(═O)—O—(C₁-C₆ alkyl),—C(═O)—NR′R″, —NR′R″, or a substituted —(C₁-C₄ alkyl), wherein thesubstituted —(C₁-C₄ alkyl) is substituted with 1-3 substituentsindependently selected from halo, —OH, —OCH₃, —S(═O)₂—CH₃, or—C(═O)—CH₃.

In some embodiments of the compound of Formula I or the pharmaceuticallyacceptable salt thereof, the hydrate thereof, or the mixture thereof, R²is an unsubstituted or substituted cyclohexyl ring. In some suchembodiments, R² is a cyclohexyl group substituted with a —(C₁-C₂ alkyl)group. In still further such embodiments, R² is a cyclohexyl groupsubstituted with a methyl group. In some such embodiments, R² is a groupof formula

where the

symbol indicates the point of attachment to the rest of the molecule. Insome embodiments, R² is an unsubstituted cyclohexyl group.

In some embodiments of the compound of Formula I or the pharmaceuticallyacceptable salt thereof, the hydrate thereof, or the mixture thereof, R′and R″ are independently selected from —H or unsubstituted —(C₁-C₄alkyl).

In some embodiments of the compound of Formula I or the pharmaceuticallyacceptable salt thereof, the hydrate thereof, or the mixture thereof, R²is an unsubstituted or substituted cyclopentyl ring. In some suchembodiments, R² is a cyclopentyl group substituted with a —(C₁-C₂ alkyl)group. In still further such embodiments, R² is a cyclopentyl groupsubstituted with a methyl group. In some such embodiments, R² is anunsubstituted cyclopentyl group.

In some embodiments of the compound of Formula I or the pharmaceuticallyacceptable salt thereof, the hydrate thereof, or the mixture thereof, R¹is a group of Formula IA or IB. In some such embodiments, R¹ is a groupof Formula IA. In other such embodiments, R¹ is a group of Formula IB.

In some embodiments of the compound of Formula I or the pharmaceuticallyacceptable salt thereof, the hydrate thereof, or the mixture thereof, R¹is a group of Formula IC or ID. In some such embodiments, R¹ is a groupof Formula IC. In other such embodiments, R¹ is a group of Formula ID.

In some embodiments of the compound of Formula I or the pharmaceuticallyacceptable salt thereof, the hydrate thereof, or the mixture thereof,

R² is a C₅-C₇ cycloalkyl group that is unsubstituted or is substitutedwith 1-3 —(C₁-C₆ alkyl) groups;

R^(3a) is selected from —H, —(C₁-C₃ alkyl), or —O—(C₁-C₃ alkyl);

R^(3b) is —H;

R^(3c) is —H;

R⁴ is —H;

R⁵ is —H;

R⁶ is selected from —H, —(C₁-C₆ alkyl), —C(═O)—(C₁-C₆ alkyl), or—C(═O)—C(═O)—OH, wherein the alkyl group of the —(C₁-C₆ alkyl) and—C(═O)—(C₁-C₆ alkyl) groups is unsubstituted or is substituted with 1-3substituents independently selected from —OH, F, —S(═O)₂—(C₁-C₆ alkyl),or —O—(C₁-C₆ alkyl);

R^(7a) is —H;

R^(7b) is —H; or is absent if R¹ is a group of Formula IB or Formula ID;

R^(7c) is —H; or is absent if R¹ is a group of Formula IA or Formula IC;

R^(8a) is —H;

R^(8b) is —H;

R^(8c) is selected from —H, —OH, or unsubstituted —(C₁-C₆ alkyl);

R^(8d) is —H;

R^(8e) is —H; and

R^(8f) is —H.

In some such embodiments, R¹ is a group of Formula IA. In otherembodiments, R¹ is a group of Formula IB. In other embodiments, R¹ is agroup of Formula IC. In other embodiments, R¹ is a group of Formula ID.

In some embodiments of the compound of Formula I or the pharmaceuticallyacceptable salt thereof, the hydrate thereof, or the mixture thereof, R⁶is selected from —H, —(C₁-C₆ alkyl), —C(═O)—(C₁-C₆ alkyl),—C(═O)—C(═O)—OH, —C(═O)—NR′R″, or —S(═O)—NR′R″, wherein the alkyl groupof the —(C₁-C₆ alkyl) and —C(═O)—(C₁-C₆ alkyl) groups is unsubstitutedor is substituted with 1-3 substituents independently selected from —OH,F, —S(═O)₂—(C₁-C₆ alkyl), —O—(C₁-C₆ alkyl), —NR′R″, or —CN.

In some embodiments of the compound of Formula I or the pharmaceuticallyacceptable salt thereof, the hydrate thereof, or the mixture thereof,the compound is the glucuronide adduct.

In some embodiments of the compound of Formula I or the pharmaceuticallyacceptable salt thereof, the hydrate thereof, or the mixture thereof,the compound has the Formula IIA

R^(3a) is selected from —H, —F, or —Cl, —(C₁-C₃ alkyl), or —O—(C₁-C₃alkyl);

R^(3b) is —H, halo, —OH, —O—(C₁-C₆ alkyl), unsubstituted —(C₁-C₆ alkyl),—NR′R″, —C(═O)—(C₁-C₆ alkyl), —C(═O)—O—(C₁-C₆ alkyl), —C(═O)—NR′R″, or asubstituted —(C₁-C₆ alkyl), wherein the substituted —(C₁-C₆ alkyl) issubstituted with 1-3 substituents independently selected from halo, —OH,—OCH₃, —CN, or —NO₂;

R⁶ is selected from —H, —(C₁-C₆ alkyl), —C(═O)—(C₁-C₆ alkyl),—C(═O)—C(═O)—OH, —C(═O)—NR′R″, or —S(═O)—NR′R″, wherein the alkyl groupof the —(C₁-C₆ alkyl) and —C(═O)—(C₁-C₆ alkyl) groups is unsubstitutedor is substituted with 1-3 substituents independently selected from —OH,F, —S(═O)₂—(C₁-C₆ alkyl), —O—(C₁-C₆ alkyl), —NR′R″, or —CN; and

R^(8c) is selected from —H, —OH, unsubstituted —(C₁-C₆ alkyl), or asubstituted —(C₁-C₆ alkyl), wherein the substituted —(C₁-C₆ alkyl) issubstituted with 1-3 substituents independently selected from —OH, halo,or —O—(C₁-C₆ alkyl).

In some embodiments of the compound of Formula IIA or thepharmaceutically acceptable salt thereof, stereoisomer thereof,pharmaceutically acceptable salt of the stereoisomer, or the mixturethereof,

R^(3a) is selected from —H, —(C₁-C₃ alkyl), or —O—(C₁-C₃ alkyl);

R^(3b) is —H;

R⁶ is selected from —H, —(C₁-C₆ alkyl), —C(═O)—(C₁-C₆ alkyl), or—C(═O)—C(═O)—OH, wherein the alkyl group of the —(C₁-C₆ alkyl) and—C(═O)—(C₁-C₆ alkyl) groups is unsubstituted or is substituted with 1-3substituents independently selected from —OH, F, —S(═O)₂—(C₁-C₆ alkyl),or —O—(C₁-C₆ alkyl); and

R^(8c) is selected from —H, unsubstituted —(C₁-C₆ alkyl), or —OH.

In some embodiments of the compound of Formula IIA or thepharmaceutically acceptable salt thereof, the hydrate thereof, or themixture thereof, R^(8c) is selected from —H, —CH₃, or —OH. In some suchembodiments, R^(8c) is —H.

In some embodiments of the compound of Formula IIA or thepharmaceutically acceptable salt thereof, the hydrate thereof, or themixture thereof, R^(3b) is —H.

In some embodiments of the compound of Formula IIA or thepharmaceutically acceptable salt thereof, the hydrate thereof, or themixture thereof, R^(3a) is —H or —OCH₃. In some such embodiments, R^(3a)is —H.

In some embodiments of the compound of Formula IIA or thepharmaceutically acceptable salt thereof, the hydrate thereof, or themixture thereof, R⁶ is selected from —H, —C(═O)—CH₃, —CH₂CH₂OH,—CH₂CH₂CH₂OH, —C(═O)—CH₂OH, —C(═O)—C(═O)—OH, —CH₂CH₂CF₃, —CH₂CH₂F,—CH₂CH₂S(═O)₂—CH₃, or —CH₂CH₂OCH₃. In some embodiments, R⁶ is —H. Inother embodiments, R⁶ is selected from —C(═O)—CH₃ or —C(═O)—CH₂OH. Instill other embodiments, R⁶ is selected from —CH₂CH₂OH, —CH₂CH₂CH₂OH, or—CH₂CH₂OCH₃. In still other embodiments, R⁶ is selected from —CH₂CH₂CF₃,—CH₂CH₂F, or —CH₂CH₂S(═O)₂—CH₃. In some embodiments of any of theseembodiments, R^(3a) is —H and R^(3b) is —H. In still other suchembodiments, R^(8c) is —H.

In some embodiments of the compound of Formula I or the pharmaceuticallyacceptable salt thereof, the hydrate thereof, or the mixture thereof,the compound has the Formula IIIA

R^(3a) is selected from —H, —F, or —Cl, —(C₁-C₃ alkyl), or —O—(C₁-C₃alkyl);

R^(3b) is —H, halo, —OH, —O—(C₁-C₆ alkyl), unsubstituted —(C₁-C₆ alkyl),—NR′R″, —C(═O)—(C₁-C₆ alkyl), —C(═O)—O—(C₁-C₆ alkyl), —C(═O)—NR′R″, or asubstituted —(C₁-C₆ alkyl), wherein the substituted —(C₁-C₆ alkyl) issubstituted with 1-3 substituents independently selected from halo, —OH,—OCH₃, —CN, or —NO₂;

R⁶ is selected from —H, —(C₁-C₆ alkyl), —C(═O)—(C₁-C₆ alkyl),—C(═O)—C(═O)—OH, —C(═O)—NR′R″, or —S(═O)—NR′R″, wherein the alkyl groupof the —(C₁-C₆ alkyl) and —C(═O)—(C₁-C₆ alkyl) groups is unsubstitutedor is substituted with 1-3 substituents independently selected from —OH,F, —S(═O)₂—(C₁-C₆ alkyl), —O—(C₁-C₆ alkyl), —NR′R″, or —CN; and

R^(8c) is selected from —H, —OH, unsubstituted —(C₁-C₆ alkyl), or asubstituted —(C₁-C₆ alkyl), wherein the substituted —(C₁-C₆ alkyl) issubstituted with 1-3 substituents independently selected from —OH, halo,or —O—(C₁-C₆ alkyl).

In some embodiments of the compound of Formula IIIA or thepharmaceutically acceptable salt thereof, the hydrate thereof, or themixture thereof,

R^(3a) is selected from —H, —(C₁-C₃ alkyl), or —O—(C₁-C₃ alkyl);

R^(3b) is —H;

R⁶ is selected from —H, —(C₁-C₆ alkyl), —C(═O)—(C₁-C₆ alkyl), or—C(═O)—C(═O)—OH, wherein the alkyl group of the —(C₁-C₆ alkyl) and—C(═O)—(C₁-C₆ alkyl) groups is unsubstituted or is substituted with 1-3substituents independently selected from —OH, F, —S(═O)₂—(C₁-C₆ alkyl),or —O—(C₁-C₆ alkyl); and

R^(8c) is selected from —H, unsubstituted —(C₁-C₆ alkyl), or —OH.

In some embodiments of the compound of Formula IIIA or thepharmaceutically acceptable salt thereof, the hydrate thereof, or themixture thereof, R^(8c) is —H.

In some embodiments of the compound of Formula IIIA or thepharmaceutically acceptable salt thereof, the hydrate thereof, or themixture thereof, R^(3b) is —H.

In some embodiments of the compound of Formula IIIA or thepharmaceutically acceptable salt thereof, the hydrate thereof, or themixture thereof, R^(3a) is —H or —OCH₃. In some such embodiments, R^(3a)is —H

In some embodiments of the compound of Formula IIIA or thepharmaceutically acceptable salt thereof, the hydrate thereof, or themixture thereof, R⁶ is selected from —H, —C(═O)—CH₃, —CH₂CH₂OH,—CH₂CH₂CH₂OH, —C(═O)—CH₂OH, —C(═O)—C(═O)—OH, —CH₂CH₂CF₃, —CH₂CH₂F,—CH₂CH₂S(═O)₂—CH₃, or —CH₂CH₂OCH₃. In other embodiments, R⁶ is —H. Instill other embodiments, R⁶ is selected from —C(═O)—CH₃ or —C(═O)—CH₂OH.In still other embodiments, R⁶ is selected from —CH₂CH₂OH, —CH₂CH₂CH₂OH,or —CH₂CH₂OCH₃. In still further embodiments, R⁶ is selected from—CH₂CH₂CF₃, —CH₂CH₂F, or —CH₂CH₂S(═O)₂—CH₃. In some embodiments of anyof these embodiments, R^(3a) is —H and R^(3b) is —H. In still other suchembodiments, R^(8c) is —H.

In some embodiments of the compound of Formula, the compound is selectedfrom

or is a pharmaceutical salt or hydrate thereof. In some embodiments, thecompound is in a neutral form whereas in others it is a pharmaceuticallyacceptable salt. In some such embodiments, the salt is selected from achloride salt, a methanesulfonate salt, or a benzenesulfonate salt. Insome such embodiments, the salt is a chloride salt. In otherembodiments, the salt is a methanesulfonate salt. In still otherembodiments, the compound is a benzenesulfonate salt. In otherembodiments, the compound is a hydrate of the neutral compound or of thesalt. For example, in some embodiments, the compound may be a chloridesalt that is a monohydrate, a dihydrate, or a trihydrate.

In some embodiments of the compound of Formula I or the pharmaceuticallyacceptable salt thereof, the hydrate thereof, or the mixture thereof,the compound is in a neutral form.

In some embodiments of the compound of Formula I or the pharmaceuticallyacceptable salt thereof, the hydrate thereof, or the mixture thereof,the compound is a pharmaceutically acceptable salt. In some suchembodiments, the salt is selected from a chloride salt, amethanesulfonate salt, or a benzenesulfonate salt. In some suchembodiments, the salt is a chloride salt. In other embodiments, the saltis a methanesulfonate salt. In still other embodiments, the compound isa benzenesulfonate salt.

Also provided are pharmaceutical compositions that include at least onepharmaceutically acceptable carrier, excipient or diluent and atherapeutically effective amount of the compound, the pharmaceuticallyacceptable salt, the hydrate, or the mixture thereof according to any ofthe embodiments described herein. In some such embodiments, the compoundis present in an amount effective for the treatment of cancer. In someembodiments, the pharmaceutical composition includes at least onepharmaceutically acceptable carrier, excipient or diluent and atherapeutically effective amount of the compound in a neutral form. Inother embodiments, the pharmaceutical composition includes at least onepharmaceutically acceptable carrier, excipient or diluent and atherapeutically effective amount of the pharmaceutically acceptablesalt. In some such embodiments, the salt is a chloride salt. In otherembodiments the salt is a methanesulfonate salt. In still otherembodiments, the salt is a benzenesulfonate salt. In some of any ofthese embodiments, the pharmaceutical composition further includes atleast one second therapeutic agent. In some such embodiments, the secondtherapeutic agent is one that is used in treating cancer. In some suchembodiments, the second therapeutic agent is used in treating acutemyeloid leukemia or any of the other cancers described below. In someembodiments, the second therapeutic agent is selected from cytosinearabinoside, daunorubicin, idarubicin, doxorubicin, cyclophosphamide,etoposide, carboplatin, fludarabine, mitoxantrone, dexamethasone,rituximab, midostaurin, a granulocyte colony-stimulating factor,filgrastim, PEG-filgrastim, lenograstim, decitabine, azacitidine,paclitaxel, gemcitibine, motesanib disphosphate, panitumumab, anantibody directed against CD33, or a CD33 bispecific T-cell engagerantibody. In other embodiments, the second therapeutic agent is selectedfrom cytosine arabinoside, daunorubicin, idarubicin, doxorubicin,cyclophosphamide, etoposide, carboplatin, fludarabine, mitoxantrone,dexamethasone, rituximab, midostaurin, a granulocyte colony-stimulatingfactor, filgrastim, PEG-filgrastim, lenograstim, decitabine,azacitidine, paclitaxel, gemcitibine, motesanib disphosphate,panitumumab. In still other such embodiments, the second therapeuticagent is selected from cytosine arabinoside, daunorubicin, idarubicin,doxorubicin, cyclophosphamide, etoposide, carboplatin, fludarabine,mitoxantrone, dexamethasone, rituximab, midostaurin, decitabine,azacitidine, paclitaxel, gemcitibine, or motesanib disphosphate. In somesuch embodiments, the second therapeutic agent is cytosine arabinoside.In other embodiments, the second therapeutic agent is daunorubicin,idarubicin, or doxorubicin. In still other embodiments, the secondtherapeutic agent is azacitidine or decitabine. In some embodiments, thesecond therapeutic agent is an anthracycline. In some embodiments, thesecond therapeutic agent is an aurora kinase inhibitor such asN-(4-((3-(2-amino-4-pyrimidinyl)-2-pyridinyl)oxy)phenyl)-4-(4-methyl-2-thienyl)-1-phthalazinamineor another compound disclosed in WO 2007/087276.

Further provided are pharmaceutical compositions that include at leastone pharmaceutically acceptable carrier, and a therapeutically effectiveamount of the composition of matter of any of the embodiments describedherein in combination with at least one additional compound such as acytotoxic agent or a compound that inhibits another kinase.

In other embodiments, the invention provides a method of treatingcancer. In some such embodiments, the cancer is resistant to otheragents such as to an anthracycline therapeutic agent. Such methodstypically include administering to a subject an effective amount of thecompound, the pharmaceutically acceptable salt thereof, the hydratethereof, or the mixture thereof of any of the embodiments or apharmaceutical composition of any of the embodiments. In some suchembodiments, the cancer is selected from acute myeloid leukemia, acutelymphoblastic leukemia myelodysplastic syndrome, multiple myeloma,chronic myeloid leukemia, acute lymphocytic leukemia, chroniclymphocytic leukemia, non-Hodgkin lymphoma, another lymphoma, anothermyeloma, or another leukemia. Examples of other cancers that may betreated with the compounds of the present invention include Burkitt'slymphoma and mantle cell lymphoma. In some such embodiments, the canceris acute myeloid leukemia. In some such embodiments, the subject is ahuman patient with mutant FLT3 such as a patient with FLT3-ITD positiveacute myeloid leukemia. In some such embodiments, the method may alsoinclude determining whether the patient has FLT3-ITD positive acutemyeloid leukemia. For example, such a method might include obtaining asample from a patient and analyzing whether the sample is FLT3-ITDpositive. In other such embodiments, the subject is a human patient withwild type FLT3 acute myeloid leukemia. In another embodiment, the canceris acute lymphoblastic leukemia. In other embodiments, the cancer isselected from breast cancer, colorectal cancer, small cell lungcarcinoma, head and neck, glioblastoma, pancreatic, gastrointestinal,liver, prostate, ovarian, testicular, endometrial, bladder, melanoma,osteosarcoma, or another sarcoma. In some embodiments, the cancer isRb-positive whereas in other embodiments, the cancer is not Rb-positive.In some embodiments, the subject is a mammal, and in some embodiments,is a human cancer patient. In some such embodiments, the cancer is ahematological cancer. In other embodiments, the cancer is a solid tumor.In some embodiments, the method includes administering to a subject aneffective amount of the compound, the pharmaceutically acceptable saltthereof, the hydrate thereof, or the mixture thereof of any of theembodiments or a pharmaceutical composition of any of the embodimentswhere the patient is a human cancer patient with mutant FLT3 and in somesuch embodiments is a human cancer patient with FLT3-ITD.

As noted above, the compounds of the invention have been found to beactive in cancer cells, for example acute myeloid leukemia cells, thatare both wild type and mutant with respect to FLT3. Therefore, in someembodiments, the invention provides methods for treating cancer inpatients with wild type FLT3 whereas in other embodiments, the inventionprovides methods for treating cancer in patients with mutant FLT3. Suchmethods typically include administering to a subject such as a humancancer patient an effective amount of the compound, the pharmaceuticallyacceptable salt thereof, the hydrate thereof, or the mixture thereof ofany of the embodiments or a pharmaceutical composition of any of theembodiments. In some such embodiments, the cancer is acute myeloidleukemia. In some such embodiments, the subject is a human patient withmutant FLT3 such as a patient that tests positive for a mutant FLT3acute myeloid leukemia. Examples of FLT3 mutants include, but are notlimited to, FLT3-ITD, FLT3 with activation loop point mutations such asFLT3-D835Y, FLT3-D835H, and FLT3-D835V, FLT3-K663Q, and FLT3-N841I.Therefore, in some embodiments, the FLT3 mutant is FLT3-ITD. In otherembodiments, the FLT3 mutant is FLT3-D835Y. In still other embodiments,the FLT3 mutant is FLT3-D835H. In other embodiments, the FLT3 mutant isFLT3-D835V. In still other embodiments, the FLT3 mutant is FLT3-K663Q.In still further such embodiments, the FLT3 mutant is FLT3-N841I. Insome such embodiments, the method may include determining whether thepatient has mutant FLT3 acute myeloid leukemia. For example, such amethod might include obtaining a sample from a patient and analyzingwhether the sample tests positive for one or more FLT3 mutant. In someembodiments, the method includes administering to a subject an effectiveamount of the compound, the pharmaceutically acceptable salt thereof,the hydrate thereof, or the mixture thereof of any of the embodiments ora pharmaceutical composition of any of the embodiments where the patientis a human cancer patient with mutant FLT3 and in some such embodimentsis a human cancer patient with acute myeloid leukemia.

In some methods of treating cancer, the patient is a human cancerpatient with a solid or hematological tumor in which the tumor exhibitslow p15^(INK4B) and/or low p6^(INK4A) expression and in some suchembodiments is a human patient with acute myeloid leukemia where thetumor exhibits low p15^(INK4B) and/or low p16^(INK4A) expression. Insome embodiments, a tumor that exhibits low p15^(INK4B) and/or lowp16^(INK4A) expression is determined using an assay that measures thelevels of these proteins or mRNA or both and compares them to the levelsin a normal myeloid tumor. This method could also be used for othertumor types. A tumor that contains less of the p15^(INK4B) and/orp16^(INK4A) proteins or mRNA or both than is found in a normal cellwould be one that exhibits low p15^(INK4B) and/or p^(INK4A) expression.In some embodiments, the tumor includes less than 90%, less than 80%,less than 70%, less than 60%, or less than 50% of p15^(INK4B) and/orp16^(INK4A) than occurs in a normal cell.

Any of the methods described herein may include the use of a secondtherapeutic agent such as those described above. For example, in oneembodiment, the invention provides a method of treating cancer whichincludes administering to a subject (a) an effective amount of thecompound, the pharmaceutically acceptable salt thereof, the hydratethereof or the mixture thereof of any of the embodiments or thepharmaceutical composition of any of the embodiments; and (b) at leastone second therapeutic agent used in the treatment of cancer. In somesuch embodiments, the second therapeutic agent is used in treating acutemyeloid leukemia or any of the other cancers described herein. In someembodiments, the second therapeutic agent is selected from cytosinearabinoside, daunorubicin, idarubicin, doxorubicin, cyclophosphamide,etoposide, carboplatin, fludarabine, mitoxantrone, dexamethasone,rituximab, midostaurin, a granulocyte colony-stimulating factor,filgrastim, PEG-filgrastim, lenograstim, decitabine, azacitidine,paclitaxel, gemcitibine, motesanib disphosphate, panitumumab, anantibody directed against CD33, or a CD33 bispecific T-cell engagerantibody. In other embodiments, the second therapeutic agent is selectedfrom cytosine arabinoside, daunorubicin, idarubicin, doxorubicin,cyclophosphamide, etoposide, carboplatin, fludarabine, mitoxantrone,dexamethasone, rituximab, midostaurin, a granulocyte colony-stimulatingfactor, filgrastim, PEG-filgrastim, lenograstim, decitabine,azacitidine, paclitaxel, gemcitibine, motesanib disphosphate,panitumumab. In still other such embodiments, the second therapeuticagent is selected from cytosine arabinoside, daunorubicin, idarubicin,doxorubicin, cyclophosphamide, etoposide, carboplatin, fludarabine,mitoxantrone, dexamethasone, rituximab, midostaurin, decitabine,azacitidine, paclitaxel, gemcitibine, or motesanib disphosphate. In somesuch embodiments, the second therapeutic agent is cytosine arabinoside.In other embodiments, the second therapeutic agent is daunorubicin,idarubicin, or doxorubicin. In still other embodiments, the secondtherapeutic agent is azacitidine or decitabine. In some embodiments, thesecond therapeutic agent is an anthracycline. In some embodiments, thesecond therapeutic agent is an aurora kinase inhibitor such asN-(4-((3-(2-amino-4-pyrimidinyl)-2-pyridinyl)oxy)phenyl)-4-(4-methyl-2-thienyl)-1-phthalazinamineor another compound disclosed in WO 2007/087276. In some embodiments,the compound, the pharmaceutically acceptable salt thereof, or themixture thereof of any of the embodiments or the pharmaceuticalcomposition of any of the embodiments is administered to the subjectafter the at least one second therapeutic agent is administered to thesubject. In other embodiments, the compound, the pharmaceuticallyacceptable salt thereof, or the hydrate thereof, or the mixture thereofof any of the embodiments or the pharmaceutical composition of any ofthe embodiments is administered to the subject before the at least onesecond therapeutic agent is administered to the subject. In still otherembodiments, the compound, the pharmaceutically acceptable salt thereof,the hydrate thereof, or the mixture thereof of any of the embodiments orthe pharmaceutical composition of any of the embodiments is administeredto the subject at the same time that the at least one second therapeuticagent is administered to the subject.

The invention further provides a compound for use in the preparation ofa medicament. Any of the compounds, salts, hydrates or mixtures of anyof the embodiments described herein can be used to prepare themedicament. In some embodiments, the cancer is selected from acutemyeloid leukemia, myelodysplastic syndrome, multiple myeloma, chronicmyeloid leukemia, acute lymphocytic leukemia, chronic lymphocyticleukemia, non-Hodgkin lymphoma, another lymphoma, another myeloma, oranother leukemia. In some such embodiments, the cancer is acute myeloidleukemia. In other embodiments, the cancer is selected from breastcancer, colorectal cancer, small cell lung carcinoma, head and neck,glioblastoma, pancreatic, gastrointestinal, liver, prostate, ovarian,testicular, endometrial, bladder, melanoma, osteosarcoma, or anothersarcoma. In some embodiments, the cancer is Rb-positive whereas in otherembodiments, the cancer is not Rb-positive. In some embodiments, thesubject is a human cancer patient and, in some such embodiments, thecancer is a hematological cancer.

Other objects, features and advantages of the invention will becomeapparent to those skilled in the art from the following description andclaims.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is a graph showing the dose dependent anti-tumor activityobserved after twice daily dosing (BID) with Example 5 inCrTac:NCR-Foxn1^(nu) nude mice with MOLM13 subcutaneous xenografttumors.

FIG. 2 is a graph showing the dose dependent anti-tumor activityobserved after twice daily dosing (BID) with Example 5 inCrTac:NCR-Foxn1^(nu) nude mice with Colo 205 subcutaneous xenografttumors.

FIG. 3 is an X-ray Powder Diffraction (XRPD) Spectrum of thehydrochloride salt of Example 5 showing 2Theta)(°) on the x-axis andIntensity (counts) on the y axis.

DETAILED DESCRIPTION OF THE INVENTION

Unless otherwise indicated, all numbers expressing quantities ofingredients, reaction conditions, and so forth used in the specificationand claims are to be understood as being modified in all instances bythe term “about.” Accordingly, unless indicated to the contrary, thenumerical parameters set forth in the following specification andattached claims are approximations that may vary depending upon thestandard deviation found in their respective testing measurements.

As used herein, if any variable occurs more than one time in a chemicalformula, its definition on each occurrence is independent of itsdefinition at every other occurrence. If the chemical structure andchemical name conflict, the chemical structure is determinative of theidentity of the compound. The compounds of the present disclosure maycontain one or more chiral centers and/or double bonds and therefore,may exist as stereoisomers, such as double-bond isomers (i.e., geometricisomers), enantiomers or diastereomers. Accordingly, any chemicalstructures within the scope of the specification depicted, in whole orin part, with a relative configuration encompass all possibleenantiomers and stereoisomers of the illustrated compounds including thestereoisomerically pure form (e.g., geometrically pure, enantiomericallypure or diastereomerically pure) and enantiomeric and stereoisomericmixtures. Enantiomeric and stereoisomeric mixtures can be resolved intothe component enantiomers or stereoisomers using separation techniquesor chiral synthesis techniques well known to the skilled artisan.

Certain compounds of the invention may possess asymmetric carbon atoms(optical centers) or double bonds; the racemates, enantiomers,diastereomers, geometric isomers and individual isomers are all intendedto be encompassed within the scope of the invention. Furthermore,atropisomers and mixtures thereof such as those resulting fromrestricted rotation about two aromatic or heteroaromatic rings bonded toone another are intended to be encompassed within the scope of theinvention. As noted above, various compounds of the invention maycontain one or more chiral centers, and can exist as racemic mixtures ofenantiomers, mixtures of diastereomers or enantiomerically or opticallypure compounds. This invention encompasses the use of stereomericallypure forms of such compounds, as well as the use of mixtures of thoseforms. For example, mixtures comprising equal or unequal amounts of theenantiomers of a particular compound of the invention may be used inmethods and compositions of the invention. These isomers may beasymmetrically synthesized or resolved using standard techniques such aschiral columns or chiral resolving agents. See, e.g., Jacques, J., etal., Enantiomers, Racemates and Resolutions (Wiley-Interscience, NewYork, 1981); Wilen, S. H., et al. (1997) Tetrahedron 33:2725; Eliel, E.L., Stereochemistry of Carbon Compounds (McGraw-Hill, N.Y., 1962); andWilen, S. H., Tables of Resolving Agents and Optical Resolutions p. 268(E. L. Eliel, Ed., Univ. of Notre Dame Press, Notre Dame, Ind., 1972).

As used herein and unless otherwise indicated, the term “stereoisomer”or “stereomerically pure” means one stereoisomer of a compound that issubstantially free of other stereoisomers of that compound. For example,a stereomerically pure compound having one chiral center will besubstantially free of the opposite enantiomer of the compound. Astereomerically pure compound having two chiral centers will besubstantially free of other diastereomers of the compound. A typicalstereomerically pure compound comprises greater than about 80% by weightof one stereoisomer of the compound and less than about 20% by weight ofother stereoisomers of the compound, more preferably greater than about90% by weight of one stereoisomer of the compound and less than about10% by weight of the other stereoisomers of the compound, even morepreferably greater than about 95% by weight of one stereoisomer of thecompound and less than about 5% by weight of the other stereoisomers ofthe compound, and most preferably greater than about 97% by weight ofone stereoisomer of the compound and less than about 3% by weight of theother stereoisomers of the compound. If the stereochemistry of astructure or a portion of a structure is not indicated with, forexample, bold or dashed lines, the structure or portion of the structureis to be interpreted as encompassing all stereoisomers of it. A bonddrawn with a wavy line indicates that both stereoisomers areencompassed.

As known by those skilled in the art, certain compounds of the inventionmay exist in one or more tautomeric forms. Because one chemicalstructure may only be used to represent one tautomeric form, it will beunderstood that for convenience, referral to a compound of a givenstructural formula includes tautomers of the structure represented bythe structural formula. The same is true with respect to stereoisomersunless a specific stereochemistry is shown or noted. For example, acompound of a specific formula includes all stereoisomers or mixturesthereof. Similarly, a pharmaceutically acceptable salt of the compoundincludes pharmaceutically acceptable salts of all individualstereoisomers or mixtures thereof.

As noted above, prodrugs also fall within the scope of chemicalentities, for example, ester or amide derivatives of the compounds ofFormula I. The term “prodrugs” includes any compounds that becomecompounds of Formula I when administered to a patient, e.g., uponmetabolic processing of the prodrug. Examples of prodrugs include, butare not limited to, acetate, formate, benzoate, carbomethoxy,carboethoxy and like derivatives of functional groups (such as alcohol,carboxylic acid, ether, ester, or amine groups) in the compounds ofFormula I.

The term “solvate” refers to the compound formed by the interaction of asolvent and a compound. Suitable solvates are pharmaceuticallyacceptable solvates, such as hydrates, including monohydrates andhemi-hydrates. The term hydrate refers to monohydrates, dihydrates, andtrihydrates.

The compounds of the invention may also contain unnatural proportions ofatomic isotopes at one or more of the atoms that constitute suchcompounds. For example, the compounds may be radiolabeled withradioactive isotopes, such as for example tritium (³H), iodine-125(¹²⁵I) or carbon-14 (¹⁴C). Radiolabeled compounds are useful astherapeutic or prophylactic agents, research reagents, e.g., assayreagents such as the ¹⁴C thymidine incorporation assay, and diagnosticagents, e.g., in vivo imaging agents. All isotopic variations of thecompounds of the invention, whether radioactive or not, are intended tobe encompassed within the scope of the invention. For example, if avariable is said to be H or —H, this means that the variable may also bedeuterium (D) or tritium (T).

As used herein, the terms “comprising” and “including” and other formsof these words are used herein in their open, non-limiting sense. Forexample, if a composition is said to comprise A, B, and C, then A, B,and C are in the composition, but D, E, and/or F may be in thecomposition as well.

“CDK4” refers to cyclin dependent kinase 4. Cyclin dependent kinases area family of serine/threonine kinases that play important roles incellular function and cell cycle progression. CDK4 is a catalyticsubunit of the protein kinase complex that is important for cell cycleG1 phase progression. The activity of CDK4 is restricted to the G1-Sphase, which is controlled by the regulatory subunits D-type cyclins andCDK inhibitor p16^(INK4A). CDK4 has been demonstrated to be responsiblefor the phosphorylation of retinoblastoma gene product (Rb). Mutationsin this gene as well as in its related proteins including D-typecyclins, p16^(INK4A) and Rb have all been found to be associated withtumorigenesis in a variety of cancers.

“CDK6” refers to cyclin dependent kinase 6. Cyclin dependent kinases area family of serine/threonine kinases that play important roles incellular function and cell cycle progression. CDK6 is a catalyticsubunit of a protein kinase complex important for cell cycle G1 phaseprogression and G1/S transition. The activity of CDK6 first appearsmid-G1 phase, which is controlled by the regulatory subunits includingD-type cyclins and members of INK4 family of CDK inhibitors. CDK6 kinasehas also been shown to phosphorylate, and thus regulate the activity of,tumor suppressor protein Rb.

“FLT3” refers to FMS-like receptor tyrosine kinase. FLT3 is a member ofthe class III tyrosine kinase receptor family. Structural features ofthis family include an extracellular domain containing fiveimmunoglobulin-like domains, a transmembrane domain a juxtamembranedomain and an intracellular region containing tyrosine kinase activity.Several mutations in FLT3 have been identified and shown to result inconstitutive activation of the receptor. In acute myeloid leukemia,these mutations are the most common genetic alteration associated withthe disease, making up approximately 25% of patients with AML. FLT3 hasbeen shown to phosphorylate and regulate the activity of STATS.

“FLT3-ITD” refers to FLT3 internal tandem duplication. FLT3-ITD is asomatic mutation in acute myeloid leukemia with variation in theposition, length, and number of duplications of the FLT3 gene. A patientwith FLT3-ITD positive acute myeloid leukemia is a patient with acutemyeloid leukemia in which the FLT3 gene exhibits this duplication.

The phrase “Rb-positive” refers to cells that express a functionalretinoblastoma (Rb) protein. Rb is a tumor suppressor that regulatesprogression of cells through the cell cycle at the G1-S transition.Phosphorylation of Rb regulates its activity. When Rb is in ahypophosphorylated state, it prevents cell cycle progression and allowsit to carry out its tumor suppressor function. Many cancer cells havebeen shown to contain mutated or deleted Rb.

The term “alkyl” refers to a saturated, branched or straight-chainmonovalent hydrocarbon group derived by the removal of one hydrogen atomfrom a single carbon atom of a parent alkane. Typical alkyl groupsinclude, but are not limited to, methyl (—CH₃); ethyl (—CH₂CH₃); propylssuch as propan-1-yl (—CH₂CH₂CH₃), and propan-2-yl (—CH(CH₂)₂); andbutyls such as butan-1-yl (—CH₂CH₂CH₂CH₃), butan-2-yl, —CH(CH₃)CH₂CH₃2-methyl-propan-1-yl (—CH₂CH(CH₃)₂, 2-methyl-propan-2-yl (—C(CH₃)₃), andtert-butyl (—C(CH₃)₃); and the like. In certain embodiments, an alkylgroup comprises 1 to 20 carbon atoms. In some embodiments, alkyl groupsinclude 1 to 6 carbon atoms whereas in other embodiments, alkyl groupsinclude 1 to 4 or 1 to 3 carbon atoms. In still other embodiments, analkyl group includes 1 or 2 carbon atoms. Branched chain alkyl groupsinclude at least 3 carbon atoms and typically include 3 to 7, or in someembodiments, 3 to 6 carbon atoms. An alkyl group having 1 to 6 carbonatoms may be referred to as a —(C₁-C₆)alkyl or —(C₁-C₆)alkyl group, analkyl group having 1 to 4 carbon atoms may be referred to as a—(C₁-C₄)alkyl or —(C₁-C₄) alkyl, and an alkyl group having 1 to 3 carbonatoms may be referred to as a —(C₁-C₃)alkyl or —(C₁-C₃)alkyl. The samedesignation system applies to alkyl groups with different numbers ofcarbon atoms. Alkyl groups may be substituted or may be unsubstituted.In some embodiments, alkyl groups are unsubstituted. In otherembodiments, an alkyl group may be substituted with one or moresubstituents. For example, in some embodiments, an alkyl group may besubstituted with 1, 2 or 3 substituents whereas in another embodiment,an alkyl group may, where permitted by valence, be substituted with 1 to5 substituents.

The term “alkoxy” refers to a radical —OR where R represents a straightor branched chain alkyl group as defined above. Representative examplesinclude, but are not limited to, methoxy (—OCH₃), ethoxy (—OCH₂CH₃),propoxy (—OCH₂CH₂CH₃), isopropoxy (—OCH(CH₃)₂), butoxy (—OCH₂CH₂CH₂CH₃),pentoxy(—OCH₂CH₂CH₂CH₂CH₃), t-butoxy (—OC(CH₃)₃) and the like. Typicalalkoxy groups include 1 to 10 carbon atoms, 1 to 6 carbon atoms, 1 to 4carbon atoms, 1 to 3 carbon atoms, or 1 to 2 carbon atoms in the Rgroup. Alkoxy groups that include 1 to 6 carbon atoms may be designatedas —O—(C₁-C₆ alkyl) groups. Similarly, alkoxy groups that include 1 to 3carbon atoms may be designated as —O—(C₁-C₃ alkyl) groups. Other alkoxygroups may be represented using the same methodology.

The term “carboxy” refers to the radical —C(O)OH which may alternativelybe written as —C(═O)OH, —C(═O)—OH, —COOH or —CO₂H. When the H atom of acarboxy group is removed and replaced with a bond to an alkyl group, thegroup may be written as —C(═O)—O-alkyl. Typical such groups includealkyl groups with 1 to 10 carbon atoms, 1 to 6 carbon atoms, 1 to 4carbon atoms, 1 to 3 carbon atoms, or 1 to 2 carbon atoms.—C(═O)—O-alkyl groups that include alkyl groups with 1 to 6 carbon atomsmay be designated as —C(═O)—O—(C₁-C₆ alkyl) groups. Similarly, suchgroups that include alkyl groups with 1 to 4 or 1 to 3 carbon atoms maybe respectively designated as —C(═O)—O—(C₁-C₄ alkyl) and —C(═O)—O—(C₁-C₃alkyl) groups. Other such groups may be represented using the samemethodology.

The term “carbonyl” refers to a radical —C(═O)—. Carbonyl groups may bebonded to alkyl groups and written as —C(═O)-alkyl groups where alkylhas the meaning set forth above. Typical alkyl groups in such—C(═O)-alkyl groups have 1 to 10 carbon atoms, 1 to 6 carbon atoms, 1 to4 carbon atoms, 1 to 3 carbon atoms, or 1 to 2 carbon atoms. The—C(═O)-alkyl groups with alkyl groups of 1 to 6 carbon atoms may bedesignated as —C(═O)—(C₁-C₆ alkyl) groups. Similarly, such groups wherethe alkyl groups have 1 to 4 or 1 to 3 carbon atoms may be respectivelydesignated as —C(═O)—(C₁-C₄ alkyl) and —C(═O)—(C₁-C₃ alkyl) groups.Other such groups may be represented using the same methodology.

The term “cyano” refers to the radical —CN which may also be written as—C≡N.

The term “cycloalkyl” refers to a saturated cyclic alkyl group derivedby the removal of one hydrogen atom from a single carbon atom of aparent cycloalkane. Typical cycloalkyl groups include, but are notlimited to, cyclopropane, cyclobutane, cyclopentane, cyclohexane,cycloheptane and the like. In certain embodiments, the cycloalkyl groupcan be C₃-C₁₀ cycloalkyl, such as, for example, C₃-C₆ cycloalkyl. Insome embodiments, a cycloalkyl group is a C₅-C₇ group such as acyclopentyl, cyclohexyl, or cycloheptyl group. Cycloalkyl groups may besubstituted or unsubstituted.

The term “bicyclic group” refers to a cyclic alkyl group that includestwo, three, or more rings derived by the removal of one hydrogen atomfrom a single carbon atom of a parent bicyclic cycloalkane. Typicalbicyclic groups include, but are not limited to, adamantyl, norbornyl,decalinyl, octahydro-1H-indenyl, bicyclo[2.2.2]octanyl,octahydropentalenyl, and the like. In certain embodiments, the bicyclicgroup is C₆-C₁₄ bicyclic group, a C₆-C₁₀ bicyclic group, a C₇-C₁₄bicyclic group, a C₇-C₁₀ bicyclic group, or a similar type bicyclicgroup. In some embodiments, a bicyclic group is a C₇-C₁₀ group.Cycloalkyl groups may be substituted or unsubstituted.

The term “heterocyclyl group” refers to a cycloalkyl group, except thatin a heterocyclyl group at least one ring atom is replaced by aheteroatom. Typically, heterocyclyl groups are characterized by thenumber of ring members and include 1, 2, or 3 heteroatoms independentlyselected from N, O, or S. In some embodiments, the heterocyclyl groupcan have 3 to 10 ring members, from 3 to 7 ring members, or from 5 to 7ring members. In some embodiments, a heterocyclyl group is a 5 to 7membered ring that includes 1, 2, or 3 heteroatoms independentlyselected from N, O, or S. Examples of heterocyclyl groups include, butare not limited to, aziridinyl, azetidinyl, oxetanyl, thiatanyl,tetrahydrofuranyl, tetrahydrothiophenyl, pyrrolidinyl, piperazinyl,piperidinyl, tetrahydro-2H-pyranyl, tetrahydro-2H-thiopyranyl,morpholinyl, thiomorpholinyl, azepanyl, oxepanyl, thiepanyl, and thelike. Heterocyclyl groups may be substituted or unsubstituted. Someexamples of alkyl substituted heterocycles include N-methylmorpholinyl,N-methylpiperidinyl, N-ethylpiperidinyl, N-methylpiperazinyl,N-propylpiperazinyl, 3-methylpiperidinyl, 2-methylpiperidinyl, and thelike.

The term “halo” or “halogen” refers to a fluoro (—F), chloro (—Cl),bromo (—Br), or iodo (—I) group.

The term “haloalkyl” refers to an alkyl group as defined above in whichat least one hydrogen atom is replaced with a halogen. Thus, the term“haloalkyl” includes “monohaloalkyl” (an alkyl substituted with onehalogen atom), “dihaloalkyl” (an alkyl substituted with two halogenatoms which may be the same or different), and “trihaloalkyl” (an alkylsubstituted with three halogen atoms which may be the same ordifferent). The term “polyhaloalkyl” refers to an alkyl group that issubstituted with two or more halogen atoms. The term “perhaloalkyl”means, unless otherwise stated, an alkyl group in which each of thehydrogen atoms is replaced with a halogen atom. For example, the term“perhaloalkyl”, includes, but is not limited to, trifluoromethyl (—CF₃),pentachloroethyl, 1,1,1-trifluoro-2-bromo-2-chloroethyl, and the like.

The term “hydroxy” refers to the hydroxyl group (—OH).

The term “nitro” refers to a radical of formula —NO₂.

The term “sulfonyl” refers to a radical —S(═O)₂—, or alternatively—SO₂—. Sulfonyl groups are typically bonded to R groups and may bewritten as —S(═O)₂—R or as —SO₂—R where R is a substituted orunsubstituted alkyl, cycloalkyl, or other specified group as definedherein. Representative examples where R is a straight chain alkyl, analkylsulfonyl, include, but are not limited to, methylsulfonyl(—S(═O)₂—CH₃), ethylsulfonyl (—S(═O)₂—CH₂CH₃), propylsulfonyl(—S(═O)₂—CH₂CH₂CH₃), butylsulfonyl(—S(═O)₂—CH₂CH₂CH₂CH₃), and the like.Typical alkylsulfonyl groups include 1 to 10 carbon atoms, 1 to 6 carbonatoms, 1 to 4 carbon atoms, 1 to 3 carbon atoms, or 1 to 2 carbon atomsin the alkyl R group. Alkylsulfonyl groups that include 1 to 6 carbonatoms may be designated as —S(═O)₂—(C₁-C₆ alkyl) groups. Similarly,alkylsulfonyl groups that include 1 to 3 carbon atoms may be designatedas —S(═O)₂—(C₁-C₃ alkyl) groups. Other alkylsulfonyl groups may bedescribed using the same methodology.

The term “amino” refers to a radical —NR′R″ where R′ and R″ areindependently chosen from —H, or substituted or unsubstituted straightor branched chain alkyl, cycloalkyl, or other specified group as definedherein. When R′ and R″ are both —H, the —NR′R″ group is a —NH₂ group.When one of R′ and R″ is —H and the other is an alkyl group, the —NR′R″is a —NH-alkyl which may also be designated as a —NH(alkyl) or—N(H)(alkyl) group. When R′ and R″ are both alkyl groups, the alkylgroups may be different, and the group may be designated as a—N(alkyl)(alkyl) group. If R′ and R″ are both alkyl groups and the alkylgroups are the same, the group may be referred to as a —N(alkyl)₂. Thealkyl groups of the R′ and R″ may be designated based on the number ofcarbon atoms in the alkyl group. For example, an R′ or R″ alkyl groupwith 1 to 6 carbon atoms may be designated as a —(C₁-C₆ alkyl).Similarly, an R′ or R″ alkyl group with 1 to 4 carbon atoms may bedesignated as a —(C₁-C₄ alkyl). By way of nonlimiting example, a —NR′R″group in which one of R′ and R″ is a —H and the other is an alkyl groupswith 1-4 carbon atoms, may be referred to as a —NH(C₁-C₄ alkyl) group oras a —N(H)(C₁-C₄ alkyl) group. Similar methodology may be used todescribe different —NR′R″ groups. Typical R′ and R″ alkyl groups of—NR′R″ groups include 1 to 10 carbon atoms, 1 to 6 carbon atoms, 1 to 4carbon atoms, 1 to 3 carbon atoms, or 1 to 2 carbon atoms.

The term “carboxamide” as used herein, refers to a group of formula—C(═O)—NR′R″ which may also be referred to a —C(═O)NR′R″ where R′ and R″are independently chosen from —H, or substituted or unsubstitutedstraight or branched chain alkyl, cycloalkyl, or other specified groupas defined herein. When R′ and R″ are both —H, the carboxamide may bewritten as a —C(═O)NH₂ or —C(═O)—NH₂ group. When one of R¹ and R″ is —Hand the other is an alkyl group, the carboxamide is a —C(═O)—NH-alkylwhich may also be designated as a —C(═O)—N(H)-alkyl, —C(═O)—N(H)(alkyl),—C(═O)N(H)-alkyl, or —C(═O)N(H)(alkyl) group. When R′ and R″ are bothalkyl groups, the alkyl groups may be different, and the group may bedesignated as a —C(═O)—N(alkyl)(alkyl) group or as a—C(═O)N(alkyl)(alkyl) group. If R′ and R″ are both alkyl groups and thealkyl groups are the same, the group may be referred to as a—C(═O)—N(alkyl)₂, or as a —C(═O)N(alkyl)₂ group. The alkyl groups of theR′ and R″groups may be designated based on the number of carbon atoms inthe alkyl group. For example, an R′ or R″ alkyl group with 1 to 6 carbonatoms may be designated as a —(C₁-C₆ alkyl). Similarly, an R′ or R″alkyl group with 1 to 4 carbon atoms may be designated as a —(C₁-C₄alkyl). By way of nonlimiting example, a —C(═O)—NR′R″ group in which oneof R′ and R″ is a —H and the other is an alkyl groups with 1-4 carbonatoms, may be referred to as a —C(═O)—NH(C₁-C₄ alkyl) group or as a—C(═O)—N(H)(C₁-C₄ alkyl) group. Similar methodology may be used todescribe different —C(═O)—NR′R″ groups. Typical R′ and R″ alkyl groupsinclude 1 to 10 carbon atoms, 1 to 6 carbon atoms, 1 to 4 carbon atoms,1 to 3 carbon atoms, or 1 to 2 carbon atoms.

The term “sulfonamide” as used herein, refers to a group of formula—S(═O)₂—NR′R″ which may also be referred to a —S(═O)₂NR′R″ where R′ andR″ are independently chosen from —H, or substituted or unsubstitutedstraight or branched chain alkyl, cycloalkyl, or other specified groupas defined herein. When R′ and R″ are both —H, the sulfonamide may bewritten as a —S(═O)₂NH₂ or —S(═O)₂—NH₂ group. When one of R′ and R″ is—H and the other is an alkyl group, the sulfonamide is a—S(═O)₂—NH-alkyl which may also be designated as a —S(═O)₂—N(H)-alkyl,—S(═O)₂—N(H)(alkyl), —S(═O)₂N(H)-alkyl, or —S(═O)₂N(H)(alkyl) group.When R′ and R″ are both alkyl groups, the alkyl groups may be different,and the group may be designated as a —S(═O)₂—N(alkyl)(alkyl) group or asa —S(═O)₂N(alkyl)(alkyl) group. If R′ and R″ are both alkyl groups andthe alkyl groups are the same, the group may be referred to as a—S(═O)₂—N(alkyl)₂, or as a —S(═O)₂N(alkyl)₂ group. The alkyl groups ofthe R′ and R″ may be designated based on the number of carbon atoms inthe alkyl group. For example, an R′ or R″ alkyl group with 1 to 6 carbonatoms may be designated as a —(C₁-C₆ alkyl). Similarly, an R′ or R″alkyl group with 1 to 4 carbon atoms may be designated as a —(C₁-C₄alkyl). By way of nonlimiting example, a —S(═O)₂—NR′R″ group in whichone of R′ and R″ is a —H and the other is an alkyl groups with 1-4carbon atoms, may be referred to as a —S(═O)₂—NH(C₁-C₄ alkyl) group oras a —S(═O)₂—N(H)(C₁-C₄ alkyl) group. Similar methodology may be used todescribe different —S(═O)₂—NR′R″ groups. Typical R′ and R″ alkyl groupsinclude 1 to 10 carbon atoms, 1 to 6 carbon atoms, 1 to 4 carbon atoms,1 to 3 carbon atoms, or 1 to 2 carbon atoms.

“Disease” refers to any disease, disorder, condition, symptom, orindication.

“Pharmaceutically acceptable” refers to generally recognized for use inanimals, and more particularly in humans.

“Pharmaceutically acceptable salt” refers to a salt of a compound thatis pharmaceutically acceptable and that possesses the desiredpharmacological activity of the parent compound. Such salts include: (1)acid addition salts, formed with inorganic acids such as hydrochloricacid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, andthe like; or formed with organic acids such as acetic acid, propionicacid, hexanoic acid, cyclopentanepropionic acid, glycolic acid, pyruvicacid, lactic acid, malonic acid, succinic acid, malic acid, maleic acid,fumaric acid, tartaric acid, citric acid, salicylic acid, benzoic acid,3-(4-hydroxybenzoyl)benzoic acid, cinnamic acid, mandelic acid,methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid,p-toluenesulfonic acid, ethane disulfonic acid and the like; or (2)salts formed when an acidic proton present in the parent compound eitheris replaced by a metal ion, e.g., an alkali metal ion, an alkaline earthion, or an aluminum ion; or coordinates with an organic base such asethanolamine, diethanolamine, triethanolamine, N-methylglucamine,dicyclohexylamine, and the like. Several salt forms may exist ashydrates such that the use of the term salt is generally defined toinclude hydrated and non-hydrated forms of the salt.

“Pharmaceutically acceptable excipient,” “pharmaceutically acceptablecarrier,” or “pharmaceutically acceptable adjuvant” refer, respectively,to an excipient, carrier or adjuvant with which at least one compound ofthe present disclosure is administered. “Pharmaceutically acceptablevehicle” refers to any of a diluent, adjuvant, excipient or carrier withwhich at least one compound of the present disclosure is administered.

“Stereoisomer” refers to an isomer that differs in the arrangement ofthe constituent atoms in space. Stereoisomers that are mirror images ofeach other and optically active are termed “enantiomers,” andstereoisomers that are not mirror images of one another and areoptically active are termed “diastereomers.”

“Subject” includes mammals and humans. The terms “human” and “subject”are used interchangeably herein.

“Therapeutically effective amount” refers to the amount of a compoundthat, when administered to a subject for treating a disease, or at leastone of the clinical symptoms of a disease or disorder, is sufficient toaffect such treatment for the disease, disorder, or symptom. The“therapeutically effective amount” can vary depending on the compound,the disease, disorder, and/or symptoms of the disease or disorder,severity of the disease, disorder, and/or symptoms of the disease ordisorder, the age of the subject to be treated, and/or the weight of thesubject to be treated. An appropriate amount in any given instance canbe readily apparent to those skilled in the art or capable ofdetermination by routine experimentation.

“Treating” or “treatment” of any disease or disorder refers to arrestingor ameliorating a disease, disorder, or at least one of the clinicalsymptoms of a disease or disorder, reducing the risk of acquiring adisease, disorder, or at least one of the clinical symptoms of a diseaseor disorder, reducing the development of a disease, disorder or at leastone of the clinical symptoms of the disease or disorder, or reducing therisk of developing a disease or disorder or at least one of the clinicalsymptoms of a disease or disorder. “Treating” or “treatment” also refersto inhibiting the disease or disorder, either physically, (e.g.,stabilization of a discernible symptom), physiologically, (e.g.,stabilization of a physical parameter), or both, or inhibiting at leastone physical parameter which may not be discernible to the subject.Further, “treating” or “treatment” refers to delaying the onset of thedisease or disorder or at least symptoms thereof in a subject which maybe exposed to or predisposed to a disease or disorder even though thatsubject does not yet experience or display symptoms of the disease ordisorder.

Reference will now be made in detail to embodiments of the presentdisclosure. While certain embodiments of the present disclosure will bedescribed, it will be understood that it is not intended to limit theembodiments of the present disclosure to those described embodiments. Tothe contrary, reference to embodiments of the present disclosure isintended to cover alternatives, modifications, and equivalents as may beincluded within the spirit and scope of the embodiments of the presentdisclosure as defined by the appended claims.

In one aspect, the invention provides a compound of Formula I:

or a pharmaceutically acceptable salt thereof, a hydrate thereof, or amixture thereof, wherein:

R¹ is a group of Formula IA, Formula IB, Formula IC, or Formula ID

wherein the

symbol indicates the point of attachment of the group of Formula IA, IB,IC, or ID to the rest of the molecule;

R² is a C₅-C₇ cycloalkyl group, is a 5 to 7-membered heterocyclyl groupthat has 1, 2, or 3 heteroatoms selected from N, O, and S, or is aC₇-C₁₀ bicyclic group; wherein the C₅-C₇ cycloalkyl group, the 5 to 7membered heterocyclyl group, or the C₇-C₁₀ bicyclic group isunsubstituted or is substituted with 1-3 substituents independentlyselected from unsubstituted —(C₁-C₆ alkyl), —OH, halo, —O—(C₁-C₆ alkyl),—CO₂H, —C(═O)—O—(C₁-C₆ alkyl), —C(═O)—NR′R″, —NR′R″, or a substituted—(C₁-C₄ alkyl), wherein the substituted —(C₁-C₄ alkyl) is substitutedwith 1-3 substituents independently selected from halo, —OH, —OCH₃,—S(═O)₂—CH₃, or —C(═O)—CH₃;

R^(3a) is selected from —H, —F, or —Cl, —(C₁-C₃ alkyl), or —O—(C₁-C₃alkyl);

R^(3b) is —H, halo, —OH, —O—(C₁-C₆ alkyl), unsubstituted —(C₁-C₆ alkyl),—NR′R″, —C(═O)—(C₁-C₆ alkyl), —C(═O)—O—(C₁-C₆ alkyl), —C(═O)—NR′R″, or asubstituted —(C₁-C₆alkyl), wherein the substituted —(C₁-C₆ alkyl) issubstituted with 1-3 substituents independently selected from halo, —OH,—OCH₃, —CN, or —NO₂;

R^(3c) is —H, —(C₁-C₃ alkyl), or halo;

R⁴ is —H;

R⁵ is —H;

R⁶ is selected from —H, —(C₁-C₆ alkyl), —C(═O)—(C₁-C₆ alkyl),—C(═O)—O—(C₁-C₆ alkyl), —C(═O)—C(═O)—OH, —C(═O)—NR′R″, or —S(═O)—NR′R″,wherein the alkyl group of the —(C₁-C₆ alkyl), —C(═O)—(C₁-C₆ alkyl), and—C(═O)—O—(C₁-C₆ alkyl) groups is unsubstituted or is substituted with1-3 substituents independently selected from —OH, F, —S(═O)₂—(C₁-C₆alkyl), —O—(C₁-C₆ alkyl), —NR′R″, or —CN;

R^(7a) is —H, —CH₃, or halo;

R^(7b) is —H, —(C₁-C₆ alkyl), or halo; or R^(7b) is absent if R¹ is agroup of Formula IB or Formula ID;

R^(7c) is —H, unsubstituted —(C₁-C₆ alkyl), halo, —O—(C₁-C₆ alkyl),—NO₂, —CN, —NR′R″, —CO₂H, —C(═O)—O—(C₁-C₆ alkyl), —C(═O)—NR′R″, or asubstituted —(C₁-C₆ alkyl), wherein the substituted —(C₁-C₆ alkyl) issubstituted with 1-3 substituents independently selected from —OH, halo,—O—(C₁-C₆ alkyl), —CN, —NR′R″, or —S(═O)₂—CH₃; or R^(7c) is absent if R¹is a group of Formula IA or Formula IC;

R^(8a) is —H, unsubstituted —(C₁-C₆ alkyl), or a substituted —(C₁-C₆alkyl), wherein the substituted —(C₁-C₆ alkyl) is substituted with 1-3substituents independently selected from —OH, halo, or —O—(C₁-C₆ alkyl);

R^(8b) is —H, unsubstituted —(C₁-C₆ alkyl), or a substituted —(C₁-C₆alkyl), wherein the substituted —(C₁-C₆ alkyl) is substituted with 1-3substituents independently selected from —OH, halo, or —O—(C₁-C₆ alkyl);or R^(8a) and R^(8b), when taken together, can represent ═O;

R^(8c) is selected from —H, —OH, unsubstituted —(C₁-C₆ alkyl), or asubstituted —(C₁-C₆ alkyl), wherein the substituted —(C₁-C₆ alkyl) issubstituted with 1-3 substituents independently selected from —OH, halo,or —O—(C₁-C₆ alkyl);

R^(8d) is —H, unsubstituted —(C₁-C₆ alkyl), or a substituted —(C₁-C₆alkyl), wherein the substituted —(C₁-C₆ alkyl) is substituted with 1-3substituents independently selected from —OH, halo, or —O—(C₁-C₆ alkyl);

R^(8e) is —H, unsubstituted —(C₁-C₆ alkyl), or a substituted —(C₁-C₆alkyl), wherein the substituted —(C₁-C₆ alkyl) is substituted with 1-3substituents independently selected from —OH, halo, or —O—(C₁-C₆ alkyl);

R^(8f) is —H, unsubstituted —(C₁-C₆ alkyl), or a substituted —(C₁-C₆alkyl), wherein the substituted —(C₁-C₆ alkyl) is substituted with 1-3substituents independently selected from —OH, halo, or —O—(C₁-C₆ alkyl);or R^(8e) and R^(8f), when taken together, can represent ═O; and

R′ and R″ are independently selected from —H, unsubstituted —(C₁-C₄alkyl), or —(C₁-C₄ alkyl) substituted with 1 to 3 substituentsindependently selected from —OH or —F.

In some embodiments of the compound of Formula I or the pharmaceuticallyacceptable salt thereof, the hydrate thereof, or the mixture thereof, R²is a C₅-C₇ cycloalkyl group that is unsubstituted or is substituted with1-3 substituents independently selected from unsubstituted —(C₁-C₆alkyl), —OH, halo, —O—(C₁-C₆ alkyl), —CO₂H, —C(═O)—O—(C₁-C₆ alkyl),—C(═O)—NR′R″, —NR′R″, or a substituted —(C₁-C₄ alkyl), wherein thesubstituted —(C₁-C₄ alkyl) is substituted with 1-3 substituentsindependently selected from halo, —OH, —OCH₃, —S(═O)₂—CH₃, or—C(═O)—CH₃.

In some embodiments of the compound of Formula I or the pharmaceuticallyacceptable salt thereof, the hydrate thereof, or the mixture thereof, R²is an unsubstituted or substituted cyclohexyl ring. In some suchembodiments, R² is a cyclohexyl group substituted with a —(C₁-C₂ alkyl)group. In still further such embodiments, R² is a cyclohexyl groupsubstituted with a methyl group. In some such embodiments, R² is a groupof formula

where the

symbol indicates the point of attachment to the rest of the molecule. Insome embodiments, R² is an unsubstituted cyclohexyl group.

In some embodiments of the compound of Formula I or the pharmaceuticallyacceptable salt thereof, the hydrate thereof, or the mixture thereof, R′and R″ are independently selected from —H or unsubstituted —(C₁-C₄alkyl).

In some embodiments of the compound of Formula I or the pharmaceuticallyacceptable salt thereof, the hydrate thereof, or the mixture thereof, R²is an unsubstituted or substituted cyclopentyl ring. In some suchembodiments, R² is a cyclopentyl group substituted with a —(C₁-C₂ alkyl)group. In still further such embodiments, R² is a cyclopentyl groupsubstituted with a methyl group. In some such embodiments, R² is anunsubstituted cyclopentyl group.

In some embodiments of the compound of Formula I or the pharmaceuticallyacceptable salt thereof, the hydrate thereof, or the mixture thereof, R¹is a group of Formula IA or IB. In some such embodiments, R¹ is a groupof Formula IA. In other such embodiments, R¹ is a group of Formula IB.

In some embodiments of the compound of Formula I or the pharmaceuticallyacceptable salt thereof, the hydrate thereof, or the mixture thereof, R¹is a group of Formula IC or ID. In some such embodiments, R¹ is a groupof Formula IC. In other such embodiments, R¹ is a group of Formula ID.

In some embodiments of the compound of Formula I or the pharmaceuticallyacceptable salt thereof, the hydrate thereof, or the mixture thereof,

R² is a C₅-C₇ cycloalkyl group that is unsubstituted or is substitutedwith 1-3 —(C₁-C₆ alkyl) groups;

R^(3a) is selected from —H, —(C₁-C₃ alkyl), or —O—(C₁-C₃ alkyl);

R^(3b) is —H;

R^(3c) is —H;

R⁴ is —H;

R⁵ is —H;

R⁶ is selected from —H, —(C₁-C₆ alkyl), —C(═O)—(C₁-C₆ alkyl), or—C(═O)—C(═O)—OH, wherein the alkyl group of the —(C₁-C₆ alkyl) and—C(═O)—(C₁-C₆ alkyl) groups is unsubstituted or is substituted with 1-3substituents independently selected from —OH, F, —S(═O)₂—(C₁-C₆ alkyl),or —O—(C₁-C₆ alkyl);

R^(7a) is —H;

R^(7b) is —H; or is absent if R¹ is a group of Formula IB or Formula ID;

R^(7c) is —H; or is absent if R¹ is a group of Formula IA or Formula IC;

R^(8a) is —H;

R^(8b) is —H;

R^(8c) is selected from —H, —OH, or unsubstituted —(C₁-C₆ alkyl);

R^(8d) is —H;

R^(8e) is —H; and

R^(8f) is —H.

In some such embodiments, R¹ is a group of Formula IA. In otherembodiments, R¹ is a group of Formula IB. In other embodiments, R¹ is agroup of Formula IC. In other embodiments, R¹ is a group of Formula ID.

In some embodiments of the compound of Formula I or the pharmaceuticallyacceptable salt thereof, the hydrate thereof, or the mixture thereof, R⁶is selected from —H, —(C₁-C₆ alkyl), —C(═O)—(C₁-C₆ alkyl),—C(═O)—C(═O)—OH, —C(═O)—NR′R″, or —S(═O)—NR′R″, wherein the alkyl groupof the —(C₁-C₆ alkyl) and —C(═O)—(C₁-C₆ alkyl) groups is unsubstitutedor is substituted with 1-3 substituents independently selected from —OH,F, —S(═O)₂—(C₁-C₆ alkyl), —O—(C₁-C₆ alkyl), —NR′R″, or —CN. In someembodiments of the compound of Formula I or the pharmaceuticallyacceptable salt thereof, the hydrate thereof, or the mixture thereof, R⁶is —C(═O)—(C₁-C₆ alkyl) and the alkyl is substituted with a —NR′R″. Insome such embodiments, R⁶ is selected from —C(═O)—CH₂—N(CH₃)₂ or—C(═O)—CH₂—N(CH₂CH₃)₂.

In some embodiments of the compound of Formula I or the pharmaceuticallyacceptable salt thereof, the hydrate thereof, or the mixture thereof,the compound is the glucuronide adduct.

In some embodiments of the compound of Formula I or the pharmaceuticallyacceptable salt thereof, the hydrate thereof, or the mixture thereof,the compound has the Formula IIA

R^(3a) is selected from —H, —F, or —Cl, —(C₁-C₃ alkyl), or —O—(C₁-C₃alkyl);

R^(3b) is —H, halo, —OH, —O—(C₁-C₆ alkyl), unsubstituted —(C₁-C₆ alkyl),—NR′R″, —C(═O)—(C₁-C₆ alkyl), —C(═O)—O—(C₁-C₆ alkyl), —C(═O)—NR′R″, or asubstituted —(C₁-C₆ alkyl), wherein the substituted —(C₁-C₆ alkyl) issubstituted with 1-3 substituents independently selected from halo, —OH,—OCH₃, —CN, or —NO₂;

R⁶ is selected from —H, —(C₁-C₆ alkyl), —C(═O)—(C₁-C₆ alkyl),—C(═O)—C(═O)—OH, —C(═O)—NR′R″, or —S(═O)—NR′R″, wherein the alkyl groupof the —(C₁-C₆ alkyl) and —C(═O)—(C₁-C₆ alkyl) groups is unsubstitutedor is substituted with 1-3 substituents independently selected from —OH,F, —S(═O)₂—(C₁-C₆ alkyl), —O—(C₁-C₆ alkyl), —NR′R″, or —CN; and

R^(8c) is selected from —H, —OH, unsubstituted —(C₁-C₆ alkyl), or asubstituted —(C₁-C₆ alkyl), wherein the substituted —(C₁-C₆ alkyl) issubstituted with 1-3 substituents independently selected from —OH, halo,or —O—(C₁-C₆ alkyl).

In some embodiments of the compound of Formula IIA or thepharmaceutically acceptable salt thereof, stereoisomer thereof,pharmaceutically acceptable salt of the stereoisomer, or the mixturethereof,

R^(3a) is selected from —H, —(C₁-C₃ alkyl), or —O—(C₁-C₃ alkyl);

R^(3b) is —H;

R⁶ is selected from —H, —(C₁-C₆ alkyl), —C(═O)—(C₁-C₆ alkyl), or—C(═O)—C(═O)—OH, wherein the alkyl group of the —(C₁-C₆ alkyl) and—C(═O)—(C₁-C₆ alkyl) groups is unsubstituted or is substituted with 1-3substituents independently selected from —OH, F, —S(═O)₂—(C₁-C₆ alkyl),or —O—(C₁-C₆ alkyl); and

R^(8c) is selected from —H, unsubstituted —(C₁-C₆ alkyl), or —OH.

In some embodiments of the compound of Formula IIA or thepharmaceutically acceptable salt thereof, the hydrate thereof, or themixture thereof, R^(8c) is selected from —H, —CH₃, or —OH. In some suchembodiments, R^(8c) is —H.

In some embodiments of the compound of Formula IIA or thepharmaceutically acceptable salt thereof, the hydrate thereof, or themixture thereof, R^(3b) is —H.

In some embodiments of the compound of Formula IIA or thepharmaceutically acceptable salt thereof, the hydrate thereof, or themixture thereof, R^(3a) is —H or —OCH₃. In some such embodiments, R^(3a)is —H.

In some embodiments of the compound of Formula IIA or thepharmaceutically acceptable salt thereof, the hydrate thereof, or themixture thereof, R⁶ is selected from —H, —C(═O)—CH₃, —CH₂CH₂OH,—CH₂CH₂CH₂OH, —C(═O)—CH₂OH, —C(═O)—C(═O)—OH, —CH₂CH₂CF₃, —CH₂CH₂F,—CH₂CH₂S(═O)₂—CH₃, or —CH₂CH₂OCH₃. In some embodiments, R⁶ is —H. Inother embodiments, R⁶ is selected from —C(═O)—CH₃ or —C(═O)—CH₂OH. Instill other embodiments, R⁶ is selected from —CH₂CH₂OH, —CH₂CH₂CH₂OH, or—CH₂CH₂OCH₃. In still other embodiments, R⁶ is selected from —CH₂CH₂CF₃,—CH₂CH₂F, or —CH₂CH₂S(═O)₂—CH₃. In some embodiments of any of theseembodiments, R^(3a) is —H and R^(3b) is —H. In still other suchembodiments, R^(8c) is —H.

In some embodiments of the compound of Formula I or the pharmaceuticallyacceptable salt thereof, the hydrate thereof, or the mixture thereof,the compound has the Formula IIIA

R^(3a) is selected from —H, —F, or —Cl, —(C₁-C₃ alkyl), or —O—(C₁-C₃alkyl);

R^(3b) is —H, halo, —OH, —O—(C₁-C₆ alkyl), unsubstituted —(C₁-C₆ alkyl),—NR′R″, —C(═O)—(C₁-C₆ alkyl), —C(═O)—O—(C₁-C₆ alkyl), —C(═O)—NR′R″, or asubstituted —(C₁-C₆ alkyl), wherein the substituted —(C₁-C₆ alkyl) issubstituted with 1-3 substituents independently selected from halo, —OH,—OCH₃, —CN, or —NO₂;

R⁶ is selected from —H, —(C₁-C₆ alkyl), —C(═O)—(C₁-C₆ alkyl),—C(═O)—C(═O)—OH, —C(═O)—NR′R″, or —S(═O)—NR′R″, wherein the alkyl groupof the —(C₁-C₆ alkyl) and —C(═O)—(C₁-C₆ alkyl) groups is unsubstitutedor is substituted with 1-3 substituents independently selected from —OH,F, —S(═O)₂—(C₁-C₆ alkyl), —O—(C₁-C₆ alkyl), —NR′R″, or —CN; and

R^(8c) is selected from —H, —OH, unsubstituted —(C₁-C₆ alkyl), or asubstituted —(C₁-C₆ alkyl), wherein the substituted —(C₁-C₆ alkyl) issubstituted with 1-3 substituents independently selected from —OH, halo,or —O—(C₁-C₆ alkyl).

In some embodiments of the compound of Formula IIIA or thepharmaceutically acceptable salt thereof, the hydrate thereof, or themixture thereof,

R^(3a) is selected from —H, —(C₁-C₃ alkyl), or —O—(C₁-C₃ alkyl);

R^(3b) is —H;

R⁶ is selected from —H, —(C₁-C₆ alkyl), —C(═O)—(C₁-C₆ alkyl), or—C(═O)—C(═O)—OH, wherein the alkyl group of the —(C₁-C₆ alkyl) and—C(═O)—(C₁-C₆ alkyl) groups is unsubstituted or is substituted with 1-3substituents independently selected from —OH, F, —S(═O)₂—(C₁-C₆ alkyl),or —O—(C₁-C₆ alkyl); and

R^(8c) is selected from —H, unsubstituted —(C₁-C₆ alkyl), or —OH.

In some embodiments of the compound of Formula IIIA or thepharmaceutically acceptable salt thereof, the hydrate thereof, or themixture thereof, R^(8c) is —H.

In some embodiments of the compound of Formula IIIA or thepharmaceutically acceptable salt thereof, the hydrate thereof, or themixture thereof, R^(3b) is —H.

In some embodiments of the compound of Formula IIIA or thepharmaceutically acceptable salt thereof, the hydrate thereof, or themixture thereof, R^(3a) is —H or —OCH₃. In some such embodiments, R^(3a)is —H

In some embodiments of the compound of Formula IIIA or thepharmaceutically acceptable salt thereof, the hydrate thereof, or themixture thereof, R⁶ is selected from —H, —C(═O)—CH₃, —CH₂CH₂OH,—CH₂CH₂CH₂OH, —C(═O)—CH₂OH, —C(═O)—C(═O)—OH, —CH₂CH₂CF₃, —CH₂CH₂F,—CH₂CH₂S(═O)₂—CH₃, or —CH₂CH₂OCH₃. In other embodiments, R⁶ is —H. Instill other embodiments, R⁶ is selected from —C(═O)—CH₃ or —C(═O)—CH₂OH.In still other embodiments, R⁶ is selected from —CH₂CH₂OH, —CH₂CH₂CH₂OH,or —CH₂CH₂OCH₃. In still further embodiments, R⁶ is selected from—CH₂CH₂CF₃, —CH₂CH₂F, or —CH₂CH₂S(═O)₂—CH₃. In some embodiments of anyof these embodiments, R^(3a) is —H and R^(3b) is —H. In still other suchembodiments, R^(8c) is —H.

In some embodiments of the compound of Formula, the compound is selectedfrom

or is a pharmaceutical salt or hydrate thereof. In some embodiments, thecompound is in a neutral form whereas in others it is a pharmaceuticallyacceptable salt. In some such embodiments, the salt is selected from achloride salt, a methanesulfonate salt, or a benzenesulfonate salt. Insome such embodiments, the salt is a chloride salt. In otherembodiments, the salt is a methanesulfonate salt. In still otherembodiments, the compound is a benzenesulfonate salt. In otherembodiments, the compound is a hydrate of the neutral compound or of thesalt. For example, in some embodiments, the compound may be a chloridesalt that is a monohydrate, a dihydrate, or a trihydrate.

In some embodiments of the compound of Formula I, the compound is

or is a pharmaceutically acceptable salt or a hydrate thereof. In someembodiments, the compound is in a neutral form whereas in others it is apharmaceutically acceptable salt. In some such embodiments, the salt isselected from a chloride salt, a methanesulfonate salt, or abenzenesulfonate salt. In some such embodiments, the salt is a chloridesalt. In other embodiments, the salt is a methanesulfonate salt. Instill other embodiments, the compound is a benzenesulfonate salt. Inother embodiments, the compound is a hydrate of the neutral compound orof the salt such as a monohydrate, a dihydrate, or a trihydrate.

In some embodiments of the compound of Formula I, the compound is

or is a pharmaceutically acceptable salt or a hydrate thereof. In someembodiments, the compound is in a neutral form whereas in others it is apharmaceutically acceptable salt. In some such embodiments, the salt isselected from a chloride salt, a methanesulfonate salt, or abenzenesulfonate salt. In some such embodiments, the salt is a chloridesalt. In other embodiments, the salt is a methanesulfonate salt. Instill other embodiments, the compound is a benzenesulfonate salt. Inother embodiments, the compound is a hydrate of the neutral compound orof the salt such as a monohydrate, a dihydrate, or a trihydrate.

In some embodiments of the compound of Formula I, the compound is

or is a pharmaceutically acceptable salt or a hydrate thereof. In someembodiments, the compound is in a neutral form whereas in others it is apharmaceutically acceptable salt. In some such embodiments, the salt isselected from a chloride salt, a methanesulfonate salt, or abenzenesulfonate salt. In some such embodiments, the salt is a chloridesalt. In other embodiments, the salt is a methanesulfonate salt. Instill other embodiments, the compound is a benzenesulfonate salt. Inother embodiments, the compound is a hydrate of the neutral compound orof the salt such as a monohydrate, a dihydrate, or a trihydrate.

In some embodiments of the compound of Formula I, the compound is

or is a pharmaceutically acceptable salt or a hydrate thereof. In someembodiments, the compound is in a neutral form whereas in others it is apharmaceutically acceptable salt. In some such embodiments, the salt isselected from a chloride salt, a methanesulfonate salt, or abenzenesulfonate salt. In some such embodiments, the salt is a chloridesalt. In other embodiments, the salt is a methanesulfonate salt. Instill other embodiments, the compound is a benzenesulfonate salt. Inother embodiments, the compound is a hydrate of the neutral compound orof the salt such as a monohydrate, a dihydrate, or a trihydrate.

In some embodiments of the compound of Formula I, the compound is

or is a pharmaceutically acceptable salt or a hydrate thereof. In someembodiments, the compound is in a neutral form whereas in others it is apharmaceutically acceptable salt. In some such embodiments, the salt isselected from a chloride salt, a methanesulfonate salt, or abenzenesulfonate salt. In some such embodiments, the salt is a chloridesalt. In other embodiments, the salt is a methanesulfonate salt. Instill other embodiments, the compound is a benzenesulfonate salt. Inother embodiments, the compound is a hydrate of the neutral compound orof the salt such as a monohydrate, a dihydrate, or a trihydrate. In somesuch embodiments, the salt is a chloride salt. In still otherembodiments, the compound is the glucuronide adduct where theglucuronide is bonded to the terminal O atom of the —C(═O)—CH₂—OH group.

In some embodiments of the compound of Formula I, the compound is

or is a pharmaceutically acceptable salt or a hydrate thereof. In someembodiments, the compound is in a neutral form whereas in others it is apharmaceutically acceptable salt. In some such embodiments, the salt isselected from a chloride salt, a methanesulfonate salt, or abenzenesulfonate salt. In some such embodiments, the salt is a chloridesalt. In other embodiments, the salt is a methanesulfonate salt. Instill other embodiments, the compound is a benzenesulfonate salt. Inother embodiments, the compound is a hydrate of the neutral compound orof the salt such as a monohydrate, a dihydrate, or a trihydrate.

In some embodiments of the compound of Formula I, the compound is

or is a pharmaceutically acceptable salt or a hydrate thereof. In someembodiments, the compound is in a neutral form whereas in others it is apharmaceutically acceptable salt. In some such embodiments, the salt isselected from a chloride salt, a methanesulfonate salt, or abenzenesulfonate salt. In some such embodiments, the salt is a chloridesalt. In other embodiments, the salt is a methanesulfonate salt. Instill other embodiments, the compound is a benzenesulfonate salt. Inother embodiments, the compound is a hydrate of the neutral compound orof the salt such as a monohydrate, a dihydrate, or a trihydrate.

In some embodiments of the compound of Formula I, the compound is

or is a pharmaceutically acceptable salt or a hydrate thereof. In someembodiments, the compound is in a neutral form whereas in others it is apharmaceutically acceptable salt. In some such embodiments, the salt isselected from a chloride salt, a methanesulfonate salt, or abenzenesulfonate salt. In some such embodiments, the salt is a chloridesalt. In other embodiments, the salt is a methanesulfonate salt. Instill other embodiments, the compound is a benzenesulfonate salt. Inother embodiments, the compound is a hydrate of the neutral compound orof the salt such as a monohydrate, a dihydrate, or a trihydrate.

In some embodiments of the compound of Formula I, the compound is

or is a pharmaceutically acceptable salt or a hydrate thereof. In someembodiments, the compound is in a neutral form whereas in others it is apharmaceutically acceptable salt. In some such embodiments, the salt isselected from a chloride salt, a methanesulfonate salt, or abenzenesulfonate salt. In some such embodiments, the salt is a chloridesalt. In other embodiments, the salt is a methanesulfonate salt. Instill other embodiments, the compound is a benzenesulfonate salt. Inother embodiments, the compound is a hydrate of the neutral compound orof the salt such as a monohydrate, a dihydrate, or a trihydrate.

In some embodiments of the compound of Formula I, the compound is

or is a pharmaceutically acceptable salt or a hydrate thereof. In someembodiments, the compound is in a neutral form whereas in others it is apharmaceutically acceptable salt. In some such embodiments, the salt isselected from a chloride salt, a methanesulfonate salt, or abenzenesulfonate salt. In some such embodiments, the salt is a chloridesalt. In other embodiments, the salt is a methanesulfonate salt. Instill other embodiments, the compound is a benzenesulfonate salt. Inother embodiments, the compound is a hydrate of the neutral compound orof the salt such as a monohydrate, a dihydrate, or a trihydrate.

In some embodiments of the compound of Formula I, the compound is

or is a pharmaceutically acceptable salt or a hydrate thereof. In someembodiments, the compound is in a neutral form whereas in others it is apharmaceutically acceptable salt. In some such embodiments, the salt isselected from a chloride salt, a methanesulfonate salt, or abenzenesulfonate salt. In some such embodiments, the salt is a chloridesalt. In other embodiments, the salt is a methanesulfonate salt. Instill other embodiments, the compound is a benzenesulfonate salt. Inother embodiments, the compound is a hydrate of the neutral compound orof the salt such as a monohydrate, a dihydrate, or a trihydrate.

In some embodiments of the compound of Formula I, the compound is

or is a pharmaceutically acceptable salt or a hydrate thereof. In someembodiments, the compound is in a neutral form whereas in others it is apharmaceutically acceptable salt. In some such embodiments, the salt isselected from a chloride salt, a methanesulfonate salt, or abenzenesulfonate salt. In some such embodiments, the salt is a chloridesalt. In other embodiments, the salt is a methanesulfonate salt. Instill other embodiments, the compound is a benzenesulfonate salt. Inother embodiments, the compound is a hydrate of the neutral compound orof the salt such as a monohydrate, a dihydrate, or a trihydrate.

In some embodiments of the compound of Formula I, the compound is

or is a pharmaceutically acceptable salt or a hydrate thereof. In someembodiments, the compound is in a neutral form whereas in others it is apharmaceutically acceptable salt. In some such embodiments, the salt isselected from a chloride salt, a methanesulfonate salt, or abenzenesulfonate salt. In some such embodiments, the salt is a chloridesalt. In other embodiments, the salt is a methanesulfonate salt. Instill other embodiments, the compound is a benzenesulfonate salt. Inother embodiments, the compound is a hydrate of the neutral compound orof the salt such as a monohydrate, a dihydrate, or a trihydrate.

In some embodiments of the compound of Formula I, the compound is

or is a pharmaceutically acceptable salt or a hydrate thereof. In someembodiments, the compound is in a neutral form whereas in others it is apharmaceutically acceptable salt. In some such embodiments, the salt isselected from a chloride salt, a methanesulfonate salt, or abenzenesulfonate salt. In some such embodiments, the salt is a chloridesalt. In other embodiments, the salt is a methanesulfonate salt. Instill other embodiments, the compound is a benzenesulfonate salt. Inother embodiments, the compound is a hydrate of the neutral compound orof the salt such as a monohydrate, a dihydrate, or a trihydrate.

In some embodiments of the compound of Formula I, the compound is

or is a pharmaceutically acceptable salt or a hydrate thereof. In someembodiments, the compound is in a neutral form whereas in others it is apharmaceutically acceptable salt. In some such embodiments, the salt isselected from a chloride salt, a methanesulfonate salt, or abenzenesulfonate salt. In some such embodiments, the salt is a chloridesalt. In other embodiments, the salt is a methanesulfonate salt. Instill other embodiments, the compound is a benzenesulfonate salt. Inother embodiments, the compound is a hydrate of the neutral compound orof the salt such as a monohydrate, a dihydrate, or a trihydrate.

In some embodiments of the compound of Formula I, the compound is

or is a pharmaceutically acceptable salt or a hydrate thereof. In someembodiments, the compound is in a neutral form whereas in others it is apharmaceutically acceptable salt. In some such embodiments, the salt isselected from a chloride salt, a methanesulfonate salt, or abenzenesulfonate salt. In some such embodiments, the salt is a chloridesalt. In other embodiments, the salt is a methanesulfonate salt. Instill other embodiments, the compound is a benzenesulfonate salt. Inother embodiments, the compound is a hydrate of the neutral compound orof the salt such as a monohydrate, a dihydrate, or a trihydrate.

In some embodiments of the compound of Formula I, the compound is

or is a pharmaceutically acceptable salt or a hydrate thereof. In someembodiments, the compound is in a neutral form whereas in others it is apharmaceutically acceptable salt. In some such embodiments, the salt isselected from a chloride salt, a methanesulfonate salt, or abenzenesulfonate salt. In some such embodiments, the salt is a chloridesalt. In other embodiments, the salt is a methanesulfonate salt. Instill other embodiments, the compound is a benzenesulfonate salt. Inother embodiments, the compound is a hydrate of the neutral compound orof the salt such as a monohydrate, a dihydrate, or a trihydrate.

In some embodiments of the compound of Formula I, the compound is

or is a pharmaceutically acceptable salt or a hydrate thereof. In someembodiments, the compound is in a neutral form whereas in others it is apharmaceutically acceptable salt. In some such embodiments, the salt isselected from a chloride salt, a methanesulfonate salt, or abenzenesulfonate salt. In some such embodiments, the salt is a chloridesalt. In other embodiments, the salt is a methanesulfonate salt. Instill other embodiments, the compound is a benzenesulfonate salt. Inother embodiments, the compound is a hydrate of the neutral compound orof the salt such as a monohydrate, a dihydrate, or a trihydrate.

In some embodiments of the compound of Formula I, the compound is

or is a pharmaceutically acceptable salt or a hydrate thereof. In someembodiments, the compound is in a neutral form whereas in others it is apharmaceutically acceptable salt. In some such embodiments, the salt isselected from a chloride salt, a methanesulfonate salt, or abenzenesulfonate salt. In some such embodiments, the salt is a chloridesalt. In other embodiments, the salt is a methanesulfonate salt. Instill other embodiments, the compound is a benzenesulfonate salt. Inother embodiments, the compound is a hydrate of the neutral compound orof the salt such as a monohydrate, a dihydrate, or a trihydrate.

In some embodiments of the compound of Formula I, the compound is

or is a pharmaceutically acceptable salt or a hydrate thereof. In someembodiments, the compound is in a neutral form whereas in others it is apharmaceutically acceptable salt. In some such embodiments, the salt isselected from a chloride salt, a methanesulfonate salt, or abenzenesulfonate salt. In some such embodiments, the salt is a chloridesalt. In other embodiments, the salt is a methanesulfonate salt. Instill other embodiments, the compound is a benzenesulfonate salt. Inother embodiments, the compound is a hydrate of the neutral compound orof the salt such as a monohydrate, a dihydrate, or a trihydrate.

In some embodiments of the compound of Formula I, the compound is

or is a pharmaceutically acceptable salt or a hydrate thereof. In someembodiments, the compound is in a neutral form whereas in others it is apharmaceutically acceptable salt. In some such embodiments, the salt isselected from a chloride salt, a methanesulfonate salt, or abenzenesulfonate salt. In some such embodiments, the salt is a chloridesalt. In other embodiments, the salt is a methanesulfonate salt. Instill other embodiments, the compound is a benzenesulfonate salt. Inother embodiments, the compound is a hydrate of the neutral compound orof the salt such as a monohydrate, a dihydrate, or a trihydrate.

In some embodiments of the compound of Formula I, the compound is

or is a pharmaceutically acceptable salt or a hydrate thereof. In someembodiments, the compound is in a neutral form whereas in others it is apharmaceutically acceptable salt. In some such embodiments, the salt isselected from a chloride salt, a methanesulfonate salt, or abenzenesulfonate salt. In some such embodiments, the salt is a chloridesalt. In other embodiments, the salt is a methanesulfonate salt. Instill other embodiments, the compound is a benzenesulfonate salt. Inother embodiments, the compound is a hydrate of the neutral compound orof the salt such as a monohydrate, a dihydrate, or a trihydrate.

In some embodiments of the compound of Formula I, the compound is

or is a pharmaceutically acceptable salt or a hydrate thereof. In someembodiments, the compound is in a neutral form whereas in others it is apharmaceutically acceptable salt. In some such embodiments, the salt isselected from a chloride salt, a methanesulfonate salt, or abenzenesulfonate salt. In some such embodiments, the salt is a chloridesalt. In other embodiments, the salt is a methanesulfonate salt. Instill other embodiments, the compound is a benzenesulfonate salt. Inother embodiments, the compound is a hydrate of the neutral compound orof the salt such as a monohydrate, a dihydrate, or a trihydrate.

In some embodiments of the compound of Formula I, the compound is

or is a pharmaceutically acceptable salt or a hydrate thereof. In someembodiments, the compound is in a neutral form whereas in others it is apharmaceutically acceptable salt. In some such embodiments, the salt isselected from a chloride salt, a methanesulfonate salt, or abenzenesulfonate salt. In some such embodiments, the salt is a chloridesalt. In other embodiments, the salt is a methanesulfonate salt. Instill other embodiments, the compound is a benzenesulfonate salt. Inother embodiments, the compound is a hydrate of the neutral compound orof the salt such as a monohydrate, a dihydrate, or a trihydrate.

In some embodiments of the compound of Formula I, the compound is

or is a pharmaceutically acceptable salt or a hydrate thereof. In someembodiments, the compound is in a neutral form whereas in others it is apharmaceutically acceptable salt. In some such embodiments, the salt isselected from a chloride salt, a methanesulfonate salt, or abenzenesulfonate salt. In some such embodiments, the salt is a chloridesalt. In other embodiments, the salt is a methanesulfonate salt. Instill other embodiments, the compound is a benzenesulfonate salt. Inother embodiments, the compound is a hydrate of the neutral compound orof the salt such as a monohydrate, a dihydrate, or a trihydrate.

In some embodiments of the compound of Formula I, the compound is

or is a pharmaceutically acceptable salt or a hydrate thereof. In someembodiments, the compound is in a neutral form whereas in others it is apharmaceutically acceptable salt. In some such embodiments, the salt isselected from a chloride salt, a methanesulfonate salt, or abenzenesulfonate salt. In some such embodiments, the salt is a chloridesalt. In other embodiments, the salt is a methanesulfonate salt. Instill other embodiments, the compound is a benzenesulfonate salt. Inother embodiments, the compound is a hydrate of the neutral compound orof the salt such as a monohydrate, a dihydrate, or a trihydrate.

In some embodiments of the compound of Formula I, the compound is

or is a pharmaceutically acceptable salt or a hydrate thereof. In someembodiments, the compound is in a neutral form whereas in others it is apharmaceutically acceptable salt. In some such embodiments, the salt isselected from a chloride salt, a methanesulfonate salt, or abenzenesulfonate salt. In some such embodiments, the salt is a chloridesalt. In other embodiments, the salt is a methanesulfonate salt. Instill other embodiments, the compound is a benzenesulfonate salt. Inother embodiments, the compound is a hydrate of the neutral compound orof the salt such as a monohydrate, a dihydrate, or a trihydrate.

In some embodiments of the compound of Formula I, the compound is

or is a pharmaceutically acceptable salt or a hydrate thereof. In someembodiments, the compound is in a neutral form whereas in others it is apharmaceutically acceptable salt. In some such embodiments, the salt isselected from a chloride salt, a methanesulfonate salt, or abenzenesulfonate salt. In some such embodiments, the salt is a chloridesalt. In other embodiments, the salt is a methanesulfonate salt. Instill other embodiments, the compound is a benzenesulfonate salt. Inother embodiments, the compound is a hydrate of the neutral compound orof the salt such as a monohydrate, a dihydrate, or a trihydrate.

In some embodiments of the compound of Formula I, the compound is

or is a pharmaceutically acceptable salt or a hydrate thereof. In someembodiments, the compound is in a neutral form whereas in others it is apharmaceutically acceptable salt. In some such embodiments, the salt isselected from a chloride salt, a methanesulfonate salt, or abenzenesulfonate salt. In some such embodiments, the salt is a chloridesalt. In other embodiments, the salt is a methanesulfonate salt. Instill other embodiments, the compound is a benzenesulfonate salt. Inother embodiments, the compound is a hydrate of the neutral compound orof the salt such as a monohydrate, a dihydrate, or a trihydrate.

In some embodiments of the compound of Formula I, the compound is

or is a pharmaceutically acceptable salt or a hydrate thereof. In someembodiments, the compound is in a neutral form whereas in others it is apharmaceutically acceptable salt. In some such embodiments, the salt isselected from a chloride salt, a methanesulfonate salt, or abenzenesulfonate salt. In some such embodiments, the salt is a chloridesalt. In other embodiments, the salt is a methanesulfonate salt. Instill other embodiments, the compound is a benzenesulfonate salt. Inother embodiments, the compound is a hydrate of the neutral compound orof the salt such as a monohydrate, a dihydrate, or a trihydrate.

In some embodiments of the compound of Formula, the compound is selectedfrom

or is a pharmaceutical salt or hydrate thereof. In some embodiments, thecompound is in a neutral form whereas in others it is a pharmaceuticallyacceptable salt. In some such embodiments, the salt is selected from achloride salt, a methanesulfonate salt, or a benzenesulfonate salt. Insome such embodiments, the salt is a chloride salt. In otherembodiments, the salt is a methanesulfonate salt. In still otherembodiments, the compound is a benzenesulfonate salt. In otherembodiments, the compound is a hydrate of the neutral compound or of thesalt such as a monohydrate, a dihydrate, or a trihydrate.

In some embodiments of the compound of Formula I or the pharmaceuticallyacceptable salt thereof, the hydrate thereof, or the mixture thereof,the compound is in a neutral form. In some such embodiments, thecompound is in a hydrate form such as a monohydrate, a dihydrate, or atrihydrate.

In some embodiments of the compound of Formula I or the pharmaceuticallyacceptable salt thereof, the hydrate thereof, or the mixture thereof,the compound is a pharmaceutically acceptable salt or a hydrate thereof.In some embodiments, the salt is selected from a chloride, citrate,tartrate, salicylate, ethanesulfonate, methanesulfonate,benzenesulfonate (besylate), tosylate, phosphate, sulfate, or ethanedisulfonate salt. In some such embodiments, the salt is selected from achloride salt, a methanesulfonate salt, or a benzenesulfonate salt. Insome embodiments, the salt is selected from a chloride, citrate,tartrate, salicylate, ethanesulfonate, benzenesulfonate, tosylate,phosphate, sulfate (½ equivalent), sulfate (1 equivalent), ethanedisulfonate, (½ equivalent), or an ethane disulfonate (1 equivalent). Insome such embodiments, the salt is selected from a chloride salt, amethanesulfonate salt, or a benzenesulfonate salt. In some suchembodiments, the salt is a chloride salt. In other embodiments, the saltis a methanesulfonate salt. In still other embodiments, the compound isa benzenesulfonate salt. In any of these embodiments, the salt may be ahydrate such as a monohydrate, a dihydrate, or a trihydrate. In someembodiments, the salt is not a hydrate.

Also provided are pharmaceutical compositions that include at least onepharmaceutically acceptable carrier, excipient or diluent and atherapeutically effective amount of the compound, the pharmaceuticallyacceptable salt, the hydrate thereof, or the mixture thereof accordingto any of the embodiments described herein. In some such embodiments,the compound is present in an amount effective for the treatment ofcancer. In some embodiments, the pharmaceutical composition includes atleast one pharmaceutically acceptable carrier, excipient or diluent anda therapeutically effective amount of the compound in a neutral formsuch as in an anhydrous form or as a hydrate. In other embodiments, thepharmaceutical composition includes at least one pharmaceuticallyacceptable carrier, excipient or diluent and a therapeutically effectiveamount of the pharmaceutically acceptable salt or a hydrate thereof. Insome such embodiments, the salt is a chloride salt. In other embodimentsthe salt is a methanesulfonate salt. In still other embodiments, thesalt is a benzenesulfonate salt. In some of any of these embodiments,the pharmaceutical composition further includes at least one secondtherapeutic agent. In some such embodiments, the second therapeuticagent is one that is used in treating cancer. In some such embodiments,the second therapeutic agent is used in treating acute myeloid leukemiaor any of the other cancers described below. In some embodiments, thesecond therapeutic agent is selected from cytosine arabinoside,daunorubicin, idarubicin, doxorubicin, cyclophosphamide, etoposide,carboplatin, fludarabine, mitoxantrone, dexamethasone, rituximab,midostaurin, a granulocyte colony-stimulating factor, filgrastim,PEG-filgrastim, lenograstim, decitabine, azacitidine, paclitaxel,gemcitibine, motesanib disphosphate, panitumumab, an antibody directedagainst CD33, or a CD33 bispecific T-cell engager antibody. In otherembodiments, the second therapeutic agent is selected from cytosinearabinoside, daunorubicin, idarubicin, doxorubicin, cyclophosphamide,etoposide, carboplatin, fludarabine, mitoxantrone, dexamethasone,rituximab, midostaurin, a granulocyte colony-stimulating factor,filgrastim, PEG-filgrastim, lenograstim, decitabine, azacitidine,paclitaxel, gemcitibine, motesanib disphosphate, panitumumab. In stillother such embodiments, the second therapeutic agent is selected fromcytosine arabinoside, daunorubicin, idarubicin, doxorubicin,cyclophosphamide, etoposide, carboplatin, fludarabine, mitoxantrone,dexamethasone, rituximab, midostaurin, decitabine, azacitidine,paclitaxel, gemcitibine, or motesanib disphosphate. In some suchembodiments, the second therapeutic agent is cytosine arabinoside. Inother embodiments, the second therapeutic agent is daunorubicin,idarubicin, or doxorubicin. In still other embodiments, the secondtherapeutic agent is azacitidine or decitabine. In some embodiments, thesecond therapeutic agent is an anthracycline. In some embodiments, thesecond therapeutic agent is an aurora kinase inhibitor such asN-(4-((3-(2-amino-4-pyrimidinyl)-2-pyridinyl)oxy)phenyl)-4-(4-methyl-2-thienyl)-1-phthalazinamineor another compound disclosed in WO 2007/087276.

Further provided are pharmaceutical compositions that include at leastone pharmaceutically acceptable carrier, and a therapeutically effectiveamount of the composition of matter of any of the embodiments describedherein in combination with at least one additional compound such as acytotoxic agent or a compound that inhibits another kinase.

In other embodiments, the invention provides a method of treatingcancer. In some such embodiments, the cancer is resistant to otheragents such as to an anthracycline therapeutic agent. Such methodstypically include administering to a subject an effective amount of thecompound, the pharmaceutically acceptable salt thereof, the hydratethereof, or the mixture thereof of any of the embodiments or apharmaceutical composition of any of the embodiments. In some suchembodiments, the cancer is selected from acute myeloid leukemia, acutelymphoblastic leukemia myelodysplastic syndrome, multiple myeloma,chronic myeloid leukemia, acute lymphocytic leukemia, chroniclymphocytic leukemia, non-Hodgkin lymphoma, another lymphoma, anothermyeloma, or another leukemia. Examples of other cancers that may betreated with the compounds of the present invention include Burkitt'slymphoma and mantle cell lymphoma. In some such embodiments, the canceris acute myeloid leukemia. In some such embodiments, the subject is apatient with acute myeloid leukemia that is 60 years of age or older. Insome such embodiments, the subject is a patient with acute myeloidleukemia that is 70 years of age or older. In some embodiments, thesubject is a human patient with mutant FLT3 such as a patient withFLT3-ITD positive acute myeloid leukemia. In some such embodiments, themethod may also include determining whether the patient has FLT3-ITDpositive acute myeloid leukemia. For example, such a method mightinclude obtaining a sample from a patient and analyzing whether thesample is FLT3-ITD positive. In other such embodiments, the subject is ahuman patient with wild type FLT3 acute myeloid leukemia. In anotherembodiment, the cancer is acute lymphoblastic leukemia. In otherembodiments, the cancer is selected from breast cancer, colorectalcancer, small cell lung carcinoma, head and neck, glioblastoma,pancreatic, gastrointestinal, liver, prostate, ovarian, testicular,endometrial, bladder, melanoma, osteosarcoma, or another sarcoma. Insome embodiments, the cancer is Rb-positive whereas in otherembodiments, the cancer is not Rb-positive. In some embodiments, thesubject is a mammal, and in some embodiments, is a human cancer patient.In some such embodiments, the cancer is a hematological cancer. In otherembodiments, the cancer is a solid tumor. In some embodiments, themethod includes administering to a subject an effective amount of thecompound, the pharmaceutically acceptable salt thereof, the hydratethereof, or the mixture thereof of any of the embodiments or apharmaceutical composition of any of the embodiments where the patientis a human cancer patient with mutant FLT3 and in some such embodimentsis a human cancer patient with FLT3-ITD. In some embodiments, the methodincludes administering to a subject an effective amount of the compound,the pharmaceutically acceptable salt thereof, the hydrate thereof, orthe mixture thereof of any of the embodiments or a pharmaceuticalcomposition of any of the embodiments where the patient is a humancancer patient with elevated levels of FLT3 ligand, for example a humanpatient with acute myeloid leukemia that has elevated FLT3 ligand.

As noted above, the compounds of the invention have been found to beactive in cancer cells, for example acute myeloid leukemia cells, thatare both wild type and mutant with respect to FLT3. Therefore, in someembodiments, the invention provides methods for treating cancer inpatients with wild type FLT3 whereas in other embodiments, the inventionprovides methods for treating cancer in patients with mutant FLT3. Suchmethods typically include administering to a subject such as a humancancer patient an effective amount of the compound, the pharmaceuticallyacceptable salt thereof, the hydrate thereof, or the mixture thereof ofany of the embodiments or a pharmaceutical composition of any of theembodiments. In some such embodiments, the cancer is acute myeloidleukemia. In some such embodiments, the subject is a human patient withmutant FLT3 such as a patient that tests positive for a mutant FLT3acute myeloid leukemia. Examples of FLT3 mutants include, but are notlimited to, FLT3-ITD, FLT3 with activation loop point mutations such asFLT3-D835Y, FLT3-D835H, and FLT3-D835V, FLT3-K663Q, and FLT3-N841I.Therefore, in some embodiments, the FLT3 mutant is FLT3-ITD. In otherembodiments, the FLT3 mutant is FLT3-D835Y. In still other embodiments,the FLT3 mutant is FLT3-D835H. In other embodiments, the FLT3 mutant isFLT3-D835V. In still other embodiments, the FLT3 mutant is FLT3-K663Q.In still further such embodiments, the FLT3 mutant is FLT3-N841I. Insome such embodiments, the method may include determining whether thepatient has mutant FLT3 acute myeloid leukemia. For example, such amethod might include obtaining a sample from a patient and analyzingwhether the sample tests positive for one or more FLT3 mutant. In someembodiments, the method includes administering to a subject an effectiveamount of the compound, the pharmaceutically acceptable salt thereof,the hydrate thereof, or the mixture thereof of any of the embodiments ora pharmaceutical composition of any of the embodiments where the patientis a human cancer patient with mutant FLT3 and in some such embodimentsis a human cancer patient with acute myeloid leukemia.

In some methods of treating cancer, the patient is a human cancerpatient with a solid or hematological tumor in which the tumor exhibitslow p 15^(INK4B) and/or low p16^(INK4A) expression and in some suchembodiments is a human patient with acute myeloid leukemia where thetumor exhibits low p15^(INK4B) and/or low p16^(INK4A) expression. Insome embodiments, a tumor that exhibits low p15^(INK4B) and/or lowp16^(INK4A) expression is determined using an assay that measures thelevels of these proteins or mRNA or both and compares them to the levelsin a normal myeloid tumor. This method could also be used for othertumor types. A tumor that contains less of the p15^(INK4B) and/orp16^(INK4A) proteins or mRNA or both than is found in a normal cellwould be one that exhibits low p15^(INK4B) and/or p16^(INK4A)expression. In some embodiments, the tumor includes less than 90%, lessthan 80%, less than 70%, less than 60%, or less than 50% of p15^(INK4B)and/or p16^(INK4A) than occurs in a normal cell such as a C34⁺ celltaken from healthy bone marrow.

Various assays may be used to evaluate whether a particular patient haswild type or mutant FLT3. For example, LabPMM (San Diego, Calif.), asubsidiary of Invivoscribe Technologies, Inc. (San Diego, Calif.) is acommercial source providing assay services such as FLT3-ITD and D835mutation assays. Such assays may be used in accordance with theinvention to screen patients or determine whether a patient, such as apatient with acute myeloid leukemia, acute lymphocytic leukemia, ormyelodysplastic syndrome has or tests positive for FLT3 mutations suchas FLT3-ITD or for mutations in the codon for the aspartic acid residueat position 835 (D835) in the FLT3 kinase domain such as in theFLT3-D835Y and FLT3-D-835H mutations. See also U.S. Pat. No. 6,846,630which is hereby incorporated by reference in its entirety and for allpurposes as if specifically set forth herein.

Assays for determining p15 mRNA expression are described by Matsuno etal. and may be used in accordance with the invention to screen or accesspatients or tumors for p15^(INK4B) expression. Matsuno et al. “p 15 mRNAexpression detected by real-time quantitative reversetranscriptase-polymerase chain reaction correlates with the methylationdensity of the gene in adult acute leukemia,” Leukemia Res., 29(5), pp.557-564 (2005) which is hereby incorporated by reference in its entiretyand for all purposes as if specifically set forth herein. The assaydescribed in this publication is a quantitative assay of p15 mRNAexpression in the bone marrow cells using real-time quantitative reversetranscriptase-polymerase chain reaction. Furthermore, this referencesdescribes the quantification of p15 mRNA expression in normal controls.

Assays for determining p16 mRNA expression are described by de Jonge etal. and may be used in accordance with the invention to screen or accesspatients or tumors for p16^(INK4A) expression. De Jonge et al. “AML atolder age: age related gene expression profiles reveal a paradoxicaldown-regulation of P16^(INK4A) mRNA with prognostic significance,”Blood, 114(14), pp. 2869-2877 (2012) which is hereby incorporated byreference in its entirety and for all purposes as if specifically setforth herein. The assay described in this publication uses quantitativereverse transcriptase polymerase chain reaction (RT-PCR). To verify thegene expression profiles, quantitative RT-PCR studies were performedusing CD34⁺ cells derived from healthy bone marrow and AML samples ofpersons of various ages. The levels of p16^(INK4B) expression wasdetermined on the isolated CD34⁺ cells derived from healthy bone marrowand derived from AML samples.

Wild type FLT3 means the protein encoded by the nucleic acid sequence ofGen Bank accession number BC144040.1. Mutant FLT3 is any FLT3 sequencethat differs from wild type FLT3 amino acid sequence(UniProtKB/Swiss-Prot Accession number P36888). Examples of FLT3 mutantsinclude, but are not limited to FLT3-ITD, FLT3-D835Y, FLT3-D835H,FLT3-D835V, FLT3-K663Q, and FLT3-N841I.

Any of the methods described herein may include the use of a secondtherapeutic agent such as those described above. For example, in oneembodiment, the invention provides a method of treating cancer whichincludes administering to a subject (a) an effective amount of thecompound, the pharmaceutically acceptable salt thereof, the hydratethereof, or the mixture thereof of any of the embodiments or thepharmaceutical composition of any of the embodiments; and (b) at leastone second therapeutic agent used in the treatment of cancer. In somesuch embodiments, the second therapeutic agent is used in treating acutemyeloid leukemia or any of the other cancers described herein. In someembodiments, the second therapeutic agent is selected from cytosinearabinoside, daunorubicin, idarubicin, doxorubicin, cyclophosphamide,etoposide, carboplatin, fludarabine, mitoxantrone, dexamethasone,rituximab, midostaurin, a granulocyte colony-stimulating factor,filgrastim, PEG-filgrastim, lenograstim, decitabine, azacitidine,paclitaxel, gemcitibine, motesanib disphosphate, panitumumab, anantibody directed against CD33, or a CD33 bispecific T-cell engagerantibody. In other embodiments, the second therapeutic agent is selectedfrom cytosine arabinoside, daunorubicin, idarubicin, doxorubicin,cyclophosphamide, etoposide, carboplatin, fludarabine, mitoxantrone,dexamethasone, rituximab, midostaurin, a granulocyte colony-stimulatingfactor, filgrastim, PEG-filgrastim, lenograstim, decitabine,azacitidine, paclitaxel, gemcitibine, motesanib disphosphate,panitumumab. In still other such embodiments, the second therapeuticagent is selected from cytosine arabinoside, daunorubicin, idarubicin,doxorubicin, cyclophosphamide, etoposide, carboplatin, fludarabine,mitoxantrone, dexamethasone, rituximab, midostaurin, decitabine,azacitidine, paclitaxel, gemcitibine, or motesanib disphosphate. In somesuch embodiments, the second therapeutic agent is cytosine arabinoside.In other embodiments, the second therapeutic agent is daunorubicin,idarubicin, or doxorubicin. In still other embodiments, the secondtherapeutic agent is azacitidine or decitabine. In some embodiments, thesecond therapeutic agent is an anthracycline. In some embodiments, thesecond therapeutic agent is an aurora kinase inhibitor such asN-(4-((3-(2-amino-4-pyrimidinyl)-2-pyridinyl)oxy)phenyl)-4-(4-methyl-2-thienyl)-1-phthalazinamineor another compound disclosed in WO 2007/087276. In some embodiments,the compound, the pharmaceutically acceptable salt thereof, the hydratethereof, or the mixture thereof of any of the embodiments or thepharmaceutical composition of any of the embodiments is administered tothe subject after the at least one second therapeutic agent isadministered to the subject. In other embodiments, the compound, thepharmaceutically acceptable salt thereof, the hydrate thereof, or themixture thereof of any of the embodiments or the pharmaceuticalcomposition of any of the embodiments is administered to the subjectbefore the at least one second therapeutic agent is administered to thesubject. In still other embodiments, the compound, the pharmaceuticallyacceptable salt thereof, the hydrate thereof, or the mixture thereof ofany of the embodiments or the pharmaceutical composition of any of theembodiments is administered to the subject at the same time that the atleast one second therapeutic agent is administered to the subject.

The invention further provides a compound for use in the preparation ofa medicament. Any of the compounds, salts, hydrates, or mixtures of anyof the embodiments described herein can be used to prepare themedicament. In some embodiments, the cancer is selected from acutemyeloid leukemia, acute lymphoblastic leukemia, myelodysplasticsyndrome, multiple myeloma, chronic myeloid leukemia, acute lymphocyticleukemia, chronic lymphocytic leukemia, non-Hodgkin lymphoma, anotherlymphoma, another myeloma, or another leukemia. In some suchembodiments, the cancer is acute myeloid leukemia. In other embodiments,the cancer is selected from breast cancer, colorectal cancer, small celllung carcinoma, head and neck, glioblastoma, pancreatic,gastrointestinal, liver, prostate, ovarian, testicular, endometrial,bladder, melanoma, osteosarcoma, or another sarcoma. In someembodiments, the cancer is Rb-positive whereas in other embodiments, thecancer is not Rb-positive. In some embodiments, the subject is a humancancer patient and, in some such embodiments, the cancer is ahematological cancer.

In another embodiment, the invention provides a method of treating aproliferation-related disorder in a mammal in need thereof. Such methodsinclude administering to the mammal a therapeutically effective amountof a compound, a salt, a hydrate, or a mixture of any of the embodimentsdescribed herein or a pharmaceutical composition comprising thecompound, salt, hydrate, or mixture. Another embodiment of the inventioncomprises treating abnormal cell growth by administering atherapeutically effective amount of a compound, salt, hydrate, ormixture of the invention or a pharmaceutical composition of theinvention to a subject in need thereof. In some embodiments, theinvention provides the use of a compound, salt, hydrate, or mixture ofany of the embodiments or a pharmaceutical composition of the inventionfor treating abnormal cell growth. The abnormal cell growth can be abenign growth or a malignant growth. In particular, the abnormal cellgrowth can be a carcinoma, sarcoma, lymphoma, or leukemia. In oneembodiment of this method, the abnormal cell growth is a cancer,including, but not limited to, lung cancer, bone cancer, pancreaticcancer, skin cancer, cancer of the head or neck, cutaneous orintraocular melanoma, uterine cancer, ovarian cancer, rectal cancer,cancer of the anal region, stomach cancer, colon cancer, breast cancer,uterine cancer, carcinoma of the fallopian tubes, carcinoma of theendometrium, carcinoma of the cervix, carcinoma of the vagina, carcinomaof the vulva, Hodgkin's Disease, cancer of the esophagus, cancer of thesmall intestine, cancer of the endocrine system, cancer of the thyroidgland, cancer of the parathyroid gland, cancer of the adrenal gland,sarcoma of soft tissue, cancer of the urethra, cancer of the penis,prostate cancer, chronic or acute leukemia, lymphocytic lymphomas,cancer of the bladder, cancer of the kidney or ureter, renal cellcarcinoma, carcinoma of the renal pelvis, neoplasms of the centralnervous system (CNS), primary CNS lymphoma, spinal axis tumors, brainstem glioma, pituitary adenoma, or a combination of one or more of theforegoing cancers. The method of the invention also comprises treating apatient having cancer where the cancer is selected from small cell lungcarcinoma, non-small cell lung carcinoma, esophageal cancer, kidneycancer, pancreatic cancer, melanoma, bladder cancer, breast cancer,colon cancer, liver cancer, lung cancer, sarcoma, stomach cancer,cholangiocarcinoma, mesothelioma, or prostate cancer. In anotherembodiment, the abnormal cell growth is a benign proliferative disease,including, but not limited to, psoriasis, benign prostatic hypertrophyor restenosis.

ADDITIONAL EMBODIMENTS

The embodiments listed below are presented in numbered form forconvenience and are in addition to the embodiments described above.

1. A compound of Formula I:

or a pharmaceutically acceptable salt thereof, a hydrate thereof, or amixture thereof, wherein:

R¹ is a group of Formula IA, Formula IB, Formula IC, or Formula ID

wherein the

symbol indicates the point of attachment of the group of Formula IA, IB,IC, or ID to the rest of the molecule;

R² is a C₅-C₇ cycloalkyl group, is a 5 to 7-membered heterocyclyl groupthat includes 1, 2, or 3 heteroatoms selected from N, O, and S, or is aC₇-C₁₀ bicyclic group; wherein the C₅-C₇ cycloalkyl group, the 5 to 7membered heterocyclyl group, or the C₇-C₁₀ bicyclic group isunsubstituted or is substituted with 1-3 substituents independentlyselected from unsubstituted —(C₁-C₆ alkyl), —OH, halo, —O—(C₁-C₆ alkyl),—CO₂H, —C(═O)—O—(C₁-C₆ alkyl), —C(═O)—NR′R″, —NR′R″, or a substituted—(C₁-C₄ alkyl), wherein the substituted —(C₁-C₄ alkyl) is substitutedwith 1-3 substituents independently selected from halo, —OH, —OCH₃,—S(═O)₂—CH₃, or —C(═O)—CH₃;

R^(3a) is selected from —H, —F, or —Cl, —(C₁-C₃ alkyl), or —O—(C₁-C₃alkyl);

R^(3b) is —H, halo, —OH, —O—(C₁-C₆ alkyl), unsubstituted —(C₁-C₆ alkyl),—NR′R″, —C(═O)—(C₁-C₆ alkyl), —C(═O)—O—(C₁-C₆ alkyl), —C(═O)—NR′R″, or asubstituted —(C₁-C₆ alkyl), wherein the substituted —(C₁-C₆ alkyl) issubstituted with 1-3 substituents independently selected from halo, —OH,—OCH₃, —CN, or —NO₂;

R^(3c) is —H, —(C₁-C₃ alkyl), or halo;

R⁴ is —H;

R⁵ is —H;

R⁶ is selected from —H, —(C₁-C₆ alkyl), —C(═O)—(C₁-C₆ alkyl),—C(═O)—O—(C₁-C₆ alkyl), —C(═O)—C(═O)—OH, —C(═O)—NR′R″, or —S(═O)—NR′R″,wherein the alkyl group of the —(C₁-C₆ alkyl), —C(═O)—(C₁-C₆ alkyl), and—C(═O)—O—(C₁-C₆ alkyl) groups is unsubstituted or is substituted with1-3 substituents independently selected from —OH, F, —S(═O)₂—(C₁-C₆alkyl), —O—(C₁-C₆ alkyl), —NR′R″, or —CN;

R^(7a) is —H, —CH₃, or halo;

R^(7b) is —H, —(C₁-C₆ alkyl), or halo; or R^(7b) is absent if R¹ is agroup of Formula IB or Formula ID;

R^(7c) is —H, unsubstituted —(C₁-C₆ alkyl), halo, —O—(C₁-C₆ alkyl),—NO₂, —CN, —NR′R″, —CO₂H, —C(═O)—O—(C₁-C₆ alkyl), —C(═O)—NR′R″, or asubstituted —(C₁-C₆ alkyl), wherein the substituted —(C₁-C₆ alkyl) issubstituted with 1-3 substituents independently selected from —OH, halo,—O—(C₁-C₆ alkyl), —CN, —NR′R″, or —S(═O)₂—CH₃; or R^(7c) is absent if R¹is a group of Formula IA or Formula IC;

R^(8a) is —H, unsubstituted —(C₁-C₆ alkyl), or a substituted —(C₁-C₆alkyl), wherein the substituted —(C₁-C₆ alkyl) is substituted with 1-3substituents independently selected from —OH, halo, or —O—(C₁-C₆ alkyl);

R^(8b) is —H, unsubstituted —(C₁-C₆ alkyl), or a substituted —(C₁-C₆alkyl), wherein the substituted —(C₁-C₆ alkyl) is substituted with 1-3substituents independently selected from —OH, halo, or —O—(C₁-C₆ alkyl);or R^(8a) and R^(8b), when taken together, can represent ═O;

R^(8c) is selected from —H, —OH, unsubstituted —(C₁-C₆ alkyl), or asubstituted —(C₁-C₆ alkyl), wherein the substituted —(C₁-C₆ alkyl) issubstituted with 1-3 substituents independently selected from —OH, halo,or —O—(C₁-C₆ alkyl);

R^(8d) is —H, unsubstituted —(C₁-C₆ alkyl), or a substituted —(C₁-C₆alkyl), wherein the substituted —(C₁-C₆ alkyl) is substituted with 1-3substituents independently selected from —OH, halo, or —O—(C₁-C₆ alkyl);

R^(8e) is —H, unsubstituted —(C₁-C₆ alkyl), or a substituted —(C₁-C₆alkyl), wherein the substituted —(C₁-C₆ alkyl) is substituted with 1-3substituents independently selected from —OH, halo, or —O—(C₁-C₆ alkyl);

R^(8f) is —H, unsubstituted —(C₁-C₆ alkyl), or a substituted —(C₁-C₆alkyl), wherein the substituted —(C₁-C₆ alkyl) is substituted with 1-3substituents independently selected from —OH, halo, or —O—(C₁-C₆ alkyl);or R^(8e) and R^(8f), when taken together, can represent ═O; and

R′ and R″ are independently selected from —H, unsubstituted —(C₁-C₄alkyl), or —(C₁-C₄ alkyl) substituted with 1 to 3 substituentsindependently selected from —OH or —F.

2. The compound of embodiment 1 or the pharmaceutically acceptable saltthereof, the hydrate thereof, or the mixture thereof, wherein R² is aC₅-C₇ cycloalkyl group that is unsubstituted or is substituted with 1-3substituents independently selected from unsubstituted —(C₁-C₆ alkyl),—OH, halo, —O—(C₁-C₆ alkyl), —CO₂H, —C(═O)—O—(C₁-C₆ alkyl),—C(═O)—NR′R″, —NR′R″, or a substituted —(C₁-C₄ alkyl), wherein thesubstituted —(C₁-C₄ alkyl) is substituted with 1-3 substituentsindependently selected from halo, —OH, —OCH₃, —S(═O)₂—CH₃, or—C(═O)—CH₃.

3. The compound of embodiment 1 or the pharmaceutically acceptable saltthereof, the hydrate thereof, or the mixture thereof, wherein R² is anunsubstituted or substituted cyclohexyl ring.

4. The compound of embodiment 3 or the pharmaceutically acceptable saltthereof, the hydrate thereof, or the mixture thereof, wherein R² is acyclohexyl group substituted with a —(C₁-C₂ alkyl) group.

5. The compound of embodiment 4 or the pharmaceutically acceptable saltthereof, the hydrate thereof, or the mixture thereof, wherein R² is agroup of formula

wherein the

symbol indicates the point of attachment to the rest of the molecule.

6. The compound of embodiment 1 or the pharmaceutically acceptable saltthereof, the hydrate thereof, or the mixture thereof, wherein R² is anunsubstituted or substituted cyclopentyl ring.

7. The compound of any one of embodiments 1-6 or the pharmaceuticallyacceptable salt thereof, the hydrate thereof, or the mixture thereof,wherein R¹ is a group of Formula IA or IB.

8. The compound of embodiment 7 or the pharmaceutically acceptable saltthereof, the hydrate thereof, or the mixture thereof, wherein R¹ is agroup of Formula IA.

9. The compound of embodiment 7 or the pharmaceutically acceptable saltthereof, the hydrate thereof, or the mixture thereof, wherein R¹ is agroup of Formula IB.

10. The compound of any one of embodiments 1-6 or the pharmaceuticallyacceptable salt thereof, the hydrate thereof, or the mixture thereof,wherein R¹ is a group of Formula IC or ID.

11. The compound of embodiment 10 or the pharmaceutically acceptablesalt thereof, the hydrate thereof, or the mixture thereof, wherein R¹ isa group of Formula IC.

12. The compound of embodiment 10 or the pharmaceutically acceptablesalt thereof, the hydrate thereof, or the mixture thereof, wherein R¹ isa group of Formula ID.

13. The compound of embodiment 1, wherein:

R² is a C₅-C₇ cycloalkyl group that is unsubstituted or is substitutedwith 1-3 —(C₁-C₆ alkyl) groups;

R^(3a) is selected from —H, —(C₁-C₃ alkyl), or —O—(C₁-C₃ alkyl);

R^(3b) is —H;

R^(3c) is —H;

R⁴ is —H;

R⁵ is —H;

R⁶ is selected from —H, —(C₁-C₆ alkyl), —C(═O)—(C₁-C₆ alkyl), or—C(═O)—C(═O)—OH, wherein the alkyl group of the —(C₁-C₆ alkyl) and—C(═O)—(C₁-C₆ alkyl) groups is unsubstituted or is substituted with 1-3substituents independently selected from —OH, F, —S(═O)₂—(C₁-C₆ alkyl),or —O—(C₁-C₆ alkyl);

R^(7a) is —H;

R^(7b) is —H; or is absent if R¹ is a group of Formula IB or Formula ID;

R^(7c) is —H; or is absent if R¹ is a group of Formula IA or Formula IC;

R^(8a) is —H;

R^(8b) is —H;

R^(8c) is selected from —H, —OH, or unsubstituted —(C₁-C₆ alkyl);

R^(8d) is —H;

R^(8e) is —H; and

R^(8f) is —H,

or the pharmaceutically acceptable salt thereof, the hydrate thereof, orthe mixture thereof.

14. The compound of embodiment 1, wherein the compound has the FormulaIIA

or the pharmaceutically acceptable salt thereof, the hydrate thereof, orthe mixture thereof,wherein:

R^(3a) is selected from —H, —F, or —Cl, —(C₁-C₃ alkyl), or —O—(C₁-C₃alkyl);

R^(3b) is —H, halo, —OH, —O—(C₁-C₆ alkyl), unsubstituted —(C₁-C₆ alkyl),—NR′R″, —C(═O)—(C₁-C₆ alkyl), —C(═O)—O—(C₁-C₆ alkyl), —C(═O)—NR′R″, or asubstituted —(C₁-C₆ alkyl), wherein the substituted —(C₁-C₆ alkyl) issubstituted with 1-3 substituents independently selected from halo, —OH,—OCH₃, —CN, or —NO₂;

R⁶ is selected from —H, —(C₁-C₆ alkyl), —C(═O)—(C₁-C₆ alkyl),—C(═O)—C(═O)—OH, —C(═O)—NR′R″, or —S(═O)—NR′R″, wherein the alkyl groupof the —(C₁-C₆ alkyl) and —C(═O)—(C₁-C₆ alkyl) groups is unsubstitutedor is substituted with 1-3 substituents independently selected from —OH,F, —S(═O)₂—(C₁-C₆ alkyl), —O—(C₁-C₆ alkyl), —NR′R″, or —CN; and

R^(8c) is selected from —H, —OH, unsubstituted —(C₁-C₆ alkyl), or asubstituted —(C₁-C₆ alkyl), wherein the substituted —(C₁-C₆ alkyl) issubstituted with 1-3 substituents independently selected from —OH, halo,or —O—(C₁-C₆ alkyl).

15. The compound of embodiment 14, wherein:

R^(3a) is selected from —H, —(C₁-C₃ alkyl), or —O—(C₁-C₃ alkyl);

R^(3b) is —H;

R⁶ is selected from —H, —(C₁-C₆ alkyl), —C(═O)—(C₁-C₆ alkyl), or—C(═O)—C(═O)—OH, wherein the alkyl group of the —(C₁-C₆ alkyl) and—C(═O)—(C₁-C₆ alkyl) groups is unsubstituted or is substituted with 1-3substituents independently selected from —OH, F, —S(═O)₂—(C₁-C₆ alkyl),or —O—(C₁-C₆ alkyl); and

R^(8c) is selected from —H, unsubstituted —(C₁-C₆ alkyl), or —OH, or thepharmaceutically acceptable salt thereof, the hydrate thereof, or themixture thereof.

16. The compound of embodiment 14 or embodiment 15 or thepharmaceutically acceptable salt thereof, the hydrate thereof, or themixture thereof, wherein R^(8c) is selected from —H, —CH₃, or —OH.

17. The compound of embodiment 16 or the pharmaceutically acceptablesalt thereof, the hydrate thereof, or the mixture thereof, whereinR^(8c) is —H.

18. The compound of any one of embodiments 14-17 or the pharmaceuticallyacceptable salt thereof, the hydrate thereof, or the mixture thereof,wherein R^(3a) is —H or —OCH₃.

19. The compound of embodiment 18 or the pharmaceutically acceptablesalt thereof, the hydrate thereof, or the mixture thereof, whereinR^(3a) is —H.

20. The compound of any one of embodiments 14-19 or the pharmaceuticallyacceptable salt thereof, the hydrate thereof, or the mixture thereof,wherein R⁶ is selected from —H, —C(═O)—CH₃, —CH₂CH₂OH, —CH₂CH₂CH₂OH,—C(═O)—CH₂OH, —C(═O)—C(═O)—OH, —CH₂CH₂CF₃, —CH₂CH₂F, —CH₂CH₂S(═O)₂—CH₃,or —CH₂CH₂OCH₃.

21. The compound of any one of embodiments 14-19 or the pharmaceuticallyacceptable salt thereof, the hydrate thereof, or the mixture thereof,wherein R⁶ is —H.

22. The compound of any one of embodiments 14-19 or the pharmaceuticallyacceptable salt thereof, the hydrate thereof, or the mixture thereof,wherein R⁶ is selected from —C(═O)—CH₃ or —C(═O)—CH₂OH.

23. The compound of any one of embodiments 14-19 or the pharmaceuticallyacceptable salt thereof, the hydrate thereof, or the mixture thereof,wherein R⁶ is selected from —CH₂CH₂OH, —CH₂CH₂CH₂OH, or —CH₂CH₂OCH₃.

24. The compound of any one of embodiments 14-19 or the pharmaceuticallyacceptable salt thereof, the hydrate thereof, or the mixture thereof,wherein R⁶ is selected from —CH₂CH₂CF₃, —CH₂CH₂F, or —CH₂CH₂S(═O)₂—CH₃.

25. The compound of any one of embodiments 14-19 or the pharmaceuticallyacceptable salt thereof, the hydrate thereof, or the mixture thereof,wherein R⁶ is —C(═O)—(C₁-C₆ alkyl) and the alkyl is substituted with a—NR′R″. 26. The compound of embodiment 25 or the pharmaceuticallyacceptable salt thereof, the hydrate thereof, or the mixture thereof,wherein R⁶ is —C(═O)—CH₂—N(CH₃)₂ or —C(═O)—CH₂—N(CH₂CH₃)₂.

27. The compound of embodiment 1, wherein the compound has the FormulaIIIA

or the pharmaceutically acceptable salt thereof, the hydrate thereof, orthe mixture thereof,wherein:

R^(3a) is selected from —H, —F, or —Cl, —(C₁-C₃ alkyl), or —O—(C₁-C₃alkyl);

R^(3b) is —H, halo, —OH, —O—(C₁-C₆ alkyl), unsubstituted —(C₁-C₆ alkyl),—NR′R —C(═O)—(C₁-C₆ alkyl), —C(═O)—O—(C₁-C₆ alkyl), —C(═O)—NR′R″, or asubstituted —(C₁-C₆ alkyl), wherein the substituted —(C₁-C₆ alkyl) issubstituted with 1-3 substituents independently selected from halo, —OH,—OCH₃, —CN, or —NO₂;

R⁶ is selected from —H, —(C₁-C₆ alkyl), —C(═O)—(C₁-C₆ alkyl),—C(═O)—C(═O)—OH, —C(═O)—NR′R″, or —S(═O)—NR′R″, wherein the alkyl groupof the —(C₁-C₆ alkyl) and —C(═O)—(C₁-C₆ alkyl) groups is unsubstitutedor is substituted with 1-3 substituents independently selected from —OH,F, —S(═O)₂—(C₁-C₆ alkyl), —O—(C₁-C₆ alkyl), —NR′R″, or —CN; and

R^(8c) is selected from —H, —OH, unsubstituted —(C₁-C₆ alkyl), or asubstituted —(C₁-C₆ alkyl), wherein the substituted —(C₁-C₆ alkyl) issubstituted with 1-3 substituents independently selected from —OH, halo,or —O—(C₁-C₆ alkyl).

28. The compound of embodiment 27, wherein:

R^(3a) is selected from —H, —(C₁-C₃ alkyl), or —O—(C₁-C₃ alkyl);

R^(3b) is —H;

R⁶ is selected from —H, —(C₁-C₆ alkyl), —C(═O)—(C₁-C₆ alkyl), or—C(═O)—C(═O)—OH, wherein the alkyl group of the —(C₁-C₆ alkyl) and—C(═O)—(C₁-C₆ alkyl) groups is unsubstituted or is substituted with 1-3substituents independently selected from —OH, F, —S(═O)₂—(C₁-C₆ alkyl),or —O—(C₁-C₆ alkyl); and

R^(8c) is selected from —H, unsubstituted —(C₁-C₆ alkyl), or —OH, or thepharmaceutically acceptable salt thereof, the hydrate thereof, or themixture thereof.

29. The compound of embodiment 27 or embodiment 28 or thepharmaceutically acceptable salt thereof, the hydrate thereof, or themixture thereof, wherein R^(8c) is —H.

30. The compound of any one of embodiments 27-29 or the pharmaceuticallyacceptable salt thereof, the hydrate thereof, or the mixture thereof,wherein R^(3a) is —H or —OCH₃.

31. The compound of embodiment 30 or the pharmaceutically acceptablesalt thereof, the hydrate thereof, or the mixture thereof, whereinR^(3a) is —H.

32. The compound of any one of embodiments 27-31 or the pharmaceuticallyacceptable salt thereof, the hydrate thereof, or the mixture thereof,wherein R⁶ is selected from —H, —C(═O)—CH₃, —CH₂CH₂OH, —CH₂CH₂CH₂OH,—C(═O)—CH₂OH, —C(═O)—C(═O)—OH, —CH₂CH₂CF₃, —CH₂CH₂F, —CH₂CH₂S(═O)₂—CH₃,or —CH₂CH₂OCH₃.

33. The compound of any one of embodiments 27-31 or the pharmaceuticallyacceptable salt thereof, the hydrate thereof, or the mixture thereof,wherein R⁶ is —H.

34. The compound of any one of embodiments 27-31 or the pharmaceuticallyacceptable salt thereof, the hydrate thereof, or the mixture thereof,wherein R⁶ is selected from —C(═O)—CH₃ or —C(═O)—CH₂OH.

35. The compound of any one of embodiments 27-31 or the pharmaceuticallyacceptable salt thereof, the hydrate thereof, or the mixture thereof,wherein R⁶ is selected from —CH₂CH₂OH, —CH₂CH₂CH₂OH, or —CH₂CH₂OCH₃.

36. The compound of any one of embodiments 27-31 or the pharmaceuticallyacceptable salt thereof, the hydrate thereof, or the mixture thereof,wherein R⁶ is selected from —CH₂CH₂CF₃, —CH₂CH₂F, or —CH₂CH₂S(═O)₂—CH₃.

37. The compound of embodiment 1, wherein the compound is selected from

or the pharmaceutically acceptable salt or the hydrate thereof.

38. The compound of embodiment 1, wherein the compound is

or the pharmaceutically acceptable salt or the hydrate thereof.

39. The compound of embodiment 1, wherein the compound is

or the pharmaceutically acceptable salt or the hydrate thereof.

40. The compound of embodiment 1, wherein the compound is

or the pharmaceutically acceptable salt or the hydrate thereof.

41. The compound of embodiment 1, wherein the compound is

or the pharmaceutically acceptable salt or the hydrate thereof.

42. The compound of embodiment 1, wherein the compound is

or the pharmaceutically acceptable salt or the hydrate thereof.

43. The compound of embodiment 1, wherein the compound is

or the pharmaceutically acceptable salt or the hydrate thereof.

44. The compound of embodiment 1, wherein the compound is

or the pharmaceutically acceptable salt or the hydrate thereof.

45. The compound of embodiment 1, wherein the compound is

or the pharmaceutically acceptable salt or the hydrate thereof.

46. The compound of embodiment 1, wherein the compound is

or the pharmaceutically acceptable salt or the hydrate thereof.

47. The compound of embodiment 1, wherein the compound is

or the pharmaceutically acceptable salt or the hydrate thereof.

48. The compound of embodiment 1, wherein the compound is

or the pharmaceutically acceptable salt or the hydrate thereof.

49. The compound of embodiment 1, wherein the compound is

or the pharmaceutically acceptable salt or the hydrate thereof.

50. The compound of embodiment 1, wherein the compound is

or the pharmaceutically acceptable salt or the hydrate thereof.

51. The compound of embodiment 1, wherein the compound is

or the pharmaceutically acceptable salt or the hydrate thereof.

52. The compound of embodiment 1, wherein the compound is

or the pharmaceutically acceptable salt or the hydrate thereof.

53. The compound of embodiment 1, wherein the compound is

or the pharmaceutically acceptable salt or the hydrate thereof.

54. The compound of embodiment 1, wherein the compound is

or the pharmaceutically acceptable salt or the hydrate thereof.

55. The compound of embodiment 1, wherein the compound is

or the pharmaceutically acceptable salt or the hydrate thereof.

56. The compound of embodiment 1, wherein the compound is

or the pharmaceutically acceptable salt or the hydrate thereof.

57. The compound of embodiment 1, wherein the compound is

or the pharmaceutically acceptable salt or the hydrate thereof.

58. The compound of embodiment 1, wherein the compound is

or the pharmaceutically acceptable salt or the hydrate thereof.

59. The compound of embodiment 1, wherein the compound is

or the pharmaceutically acceptable salt or the hydrate thereof.

60. The compound of embodiment 1, wherein the compound is

or the pharmaceutically acceptable salt or the hydrate thereof.

61. The compound of embodiment 1, wherein the compound is

or the pharmaceutically acceptable salt or the hydrate thereof.

62. The compound of embodiment 1, wherein the compound is

or the pharmaceutically acceptable salt or the hydrate thereof.

63. The compound of embodiment 1, wherein the compound is

or the pharmaceutically acceptable salt or the hydrate thereof.

64. The compound of embodiment 1, wherein the compound is

or the pharmaceutically acceptable salt or the hydrate thereof.

65. The compound of embodiment 1, wherein the compound is

or the pharmaceutically acceptable salt or the hydrate thereof.

66. The compound of embodiment 1, wherein the compound is

or the pharmaceutically acceptable salt or the hydrate thereof.

67. The compound of embodiment 1, wherein the compound is

or the pharmaceutically acceptable salt or the hydrate thereof.

68. The compound of embodiment 1, wherein the compound is selected from

or the pharmaceutically acceptable salt or the hydrate thereof.

69. The compound of embodiment 1, wherein the compound is

or the pharmaceutically acceptable salt or the hydrate thereof.

70. The compound of embodiment 1, wherein the compound is

or the pharmaceutically acceptable salt or the hydrate thereof.

71. The compound of embodiment 1, wherein the compound is

or the pharmaceutically acceptable salt or the hydrate thereof.

72. The compound of embodiment 1, wherein the compound is

or the pharmaceutically acceptable salt or the hydrate thereof.

73. The compound of embodiment 1, wherein the compound is

or the pharmaceutically acceptable salt or the hydrate thereof.

74. The compound of embodiment 1, wherein the compound is

or the pharmaceutically acceptable salt or the hydrate thereof.

75. The compound of embodiment 1, wherein the compound is

or the pharmaceutically acceptable salt or the hydrate thereof.

76. The compound of embodiment 1, wherein the compound is

or the pharmaceutically acceptable salt or the hydrate thereof.

77. The compound of any one of embodiments 1-76 in a neutral form.

78. The pharmaceutically acceptable salt of any one of embodiments 1-76.

79. The pharmaceutically acceptable salt of embodiment 78, wherein thepharmaceutically acceptable salt is selected from a chloride salt, amethanesulfonate salt, or a benzenesulfonate salt.

80. The pharmaceutically acceptable salt of embodiment 78, wherein thepharmaceutically acceptable salt is a chloride salt.

81. The pharmaceutically acceptable salt of embodiment 78, wherein thepharmaceutically acceptable salt is a methanesulfonate salt.

82. The pharmaceutically acceptable salt of embodiment 78, wherein thepharmaceutically acceptable salt is a benzenesulfonate salt.

83. A pharmaceutical composition, the pharmaceutical compositioncomprising a therapeutically effective amount of the compound, thepharmaceutically acceptable salt, the hydrate thereof, or the mixturethereof according to any one of embodiments 1-82 and at least onepharmaceutically acceptable excipient, carrier, or diluent.

84. A method of treating cancer, the method comprising: administering toa subject an effective amount of the compound, the pharmaceuticallyacceptable salt thereof, the hydrate thereof, or the mixture thereof ofany one of embodiments 1-82 or the pharmaceutical composition ofembodiment 83.

85. The method of embodiment 84, wherein the cancer is selected fromacute myeloid leukemia, acute lymphoblastic leukemia, myelodysplasticsyndrome, multiple myeloma, chronic myeloid leukemia, acute lymphocyticleukemia, chronic lymphocytic leukemia, non-Hodgkin lymphoma, anotherlymphoma, another myeloma, or another leukemia.

86. The method of embodiment 85, wherein the cancer is acute myeloidleukemia.

87. The method of embodiment 84, wherein the cancer is selected frombreast cancer, colorectal cancer, small cell lung carcinoma, head andneck, glioblastoma, pancreatic, gastrointestinal, liver, prostate,ovarian, testicular, endometrial, bladder, melanoma, osteosarcoma, oranother sarcoma.

88. The method of any one of embodiments 84-88, wherein the cancer isRb-positive.

89. The method of embodiment 84, wherein the subject is a human patient,and the cancer is a hematological cancer.

90. A method of treating cancer, the method comprising: administering toa subject

(a) an effective amount of the compound, the pharmaceutically acceptablesalt thereof, the hydrate thereof, or the mixture thereof of any one ofembodiments 1-82 or the pharmaceutical composition of embodiment 83; and

(b) at least one second therapeutic agent used in the treatment ofcancer.

91. The method of embodiment 90, wherein the second therapeutic agent iscytosine arabinoside, daunorubicin, idarubicin, doxorubicin,cyclophosphamide, etoposide, carboplatin, fludarabine, mitoxantrone,dexamethasone, rituximab, midostaurin, a granulocyte colony-stimulatingfactor, filgrastim, PEG-filgrastim, lenograstim, decitabine,azacitidine, paclitaxel, gemcitibine, motesanib disphosphate,panitumumab, an antibody directed against CD33, or a CD33 bispecificT-cell engager antibody.

92. The method of embodiment 90, wherein the second therapeutic agent iscytosine arabinoside.

93. The method of embodiment 90, wherein the second therapeutic agent isan agent used in the treatment of acute myeloid leukemia.

94. The method of any one of embodiments 90-93, wherein the effectiveamount of the compound, the pharmaceutically acceptable salt thereof,the hydrate thereof, or the mixture thereof of any one of embodiments1-82 or the pharmaceutical composition of embodiment 83 is administeredto the subject after the at least one second therapeutic agent isadministered to the subject.

95. The method of any one of embodiments 90-93, wherein the effectiveamount of the compound, the pharmaceutically acceptable salt thereof,the hydrate thereof, or the mixture thereof of any one of embodiments1-82 or the pharmaceutical composition of embodiment 83 is administeredto the subject before the at least one second therapeutic agent isadministered to the subject.

96. The method of any one of embodiments 90-93, wherein the effectiveamount of the compound, the pharmaceutically acceptable salt thereof,the hydrate thereof, or the mixture thereof of any one of embodiments1-82 or the pharmaceutical composition of embodiment 83 is administeredto the subject at the same time that the at least one second therapeuticagent is administered to the subject.

97. The compound of any one of embodiments 1-82 in the preparation of amedicament.

98. The compound of any one of embodiments 1-82 or the pharmaceuticallyacceptable salt, or the hydrate thereof, or the mixture thereof or thepharmaceutical composition of embodiment 83 for treating cancer.

99. The compound of embodiment 98 or the pharmaceutically acceptablesalt, or the hydrate thereof, or the mixture thereof, wherein the canceris selected from acute myeloid leukemia, acute lymphoblastic leukemia,myelodysplastic syndrome, multiple myeloma, chronic myeloid leukemia,acute lymphocytic leukemia, chronic lymphocytic leukemia, non-Hodgkinlymphoma, another lymphoma, another myeloma, or another leukemia.

100. The compound of embodiment 98 or the pharmaceutically acceptablesalt, or the hydrate thereof, or the mixture thereof, wherein the canceris acute myeloid leukemia.

101. The compound of embodiment 98 or the pharmaceutically acceptablesalt, or the hydrate thereof, or the mixture thereof, wherein the canceris selected from breast cancer, colorectal cancer, small cell lungcarcinoma, head and neck, glioblastoma, pancreatic, gastrointestinal,liver, prostate, ovarian, testicular, endometrial, bladder, melanoma,osteosarcoma, or another sarcoma.

102. The compound of embodiment 98 or the pharmaceutically acceptablesalt, or the hydrate thereof, or the mixture thereof, wherein the canceris Rb-positive.

103. The compound of embodiment 98 or the pharmaceutically acceptablesalt, or the hydrate thereof, or the mixture thereof, wherein thecompound is for use in treating a human cancer patient, and the canceris a hematological cancer.

104. The compound, the pharmaceutically acceptable salt thereof, thehydrate thereof, or the mixture thereof of any one of embodiments 1-82or the pharmaceutical composition of embodiment 83 for treating cancer,the treatment of cancer comprising: administering to a subject

(a) an effective amount of the compound, the pharmaceutically acceptablesalt thereof, the hydrate thereof, or the mixture thereof of any one ofembodiments 1-82 or the pharmaceutical composition of embodiment 83; and

(b) at least one second therapeutic agent used in the treatment ofcancer.

105. The compound of embodiment 104 or the pharmaceutically acceptablesalt, or the hydrate thereof, or the mixture thereof, wherein the secondtherapeutic agent is cytosine arabinoside, daunorubicin, idarubicin,doxorubicin, cyclophosphamide, etoposide, carboplatin, fludarabine,mitoxantrone, dexamethasone, rituximab, midostaurin, a granulocytecolony-stimulating factor, filgrastim, PEG-filgrastim, lenograstim,decitabine, azacitidine, paclitaxel, gemcitibine, motesanibdisphosphate, panitumumab, an antibody directed against CD33, or a CD33bispecific T-cell engager antibody.

106. The compound of embodiment 104 or the pharmaceutically acceptablesalt, or the hydrate thereof, or the mixture thereof, wherein the secondtherapeutic agent is cytosine arabinoside.

107. The compound of embodiment 104 or the pharmaceutically acceptablesalt, or the hydrate thereof, or the mixture thereof, wherein the secondtherapeutic agent is an agent used in the treatment of acute myeloidleukemia.

108. The compound of any one of embodiments 104-107 or thepharmaceutically acceptable salt, or the hydrate thereof, or the mixturethereof, wherein the effective amount of the compound, thepharmaceutically acceptable salt thereof, or the mixture thereof of anyone of embodiments 1-82 or the pharmaceutical composition of embodiment83 is administered to the subject after the at least one secondtherapeutic agent is administered to the subject.

109. The compound of any one of embodiments 104-107 or thepharmaceutically acceptable salt, or the hydrate thereof, or the mixturethereof, wherein the effective amount of the compound, thepharmaceutically acceptable salt thereof, or the mixture thereof of anyone of embodiments 1-82 or the pharmaceutical composition of embodiment83 is administered to the subject before the at least one secondtherapeutic agent is administered to the subject.

110. The compound of any one of embodiments 104-107 or thepharmaceutically acceptable salt, or the hydrate thereof, or the mixturethereof, wherein the effective amount of the compound, thepharmaceutically acceptable salt thereof, or the mixture thereof of anyone of embodiments 1-74 or the pharmaceutical composition of embodiment75 is administered to the subject at the same time that the at least onesecond therapeutic agent is administered to the subject.

The pharmaceutical compositions or compositions for the administrationof the compounds of this invention may conveniently be presented in unitdosage form and may be prepared by any of the methods well known in theart. All methods include the step of bringing the active ingredient intoassociation with the carrier which constitutes one or more accessoryingredients. In general, the pharmaceutical compositions are prepared byuniformly and intimately bringing the active ingredient into associationwith a liquid carrier or a finely divided solid carrier or both, andthen, if necessary, shaping the product into the desired formulation. Inthe pharmaceutical composition, the active object compound is includedin an amount sufficient to produce the desired effect upon the processor condition of diseases.

The pharmaceutical compositions containing the active ingredient may bein a form suitable for oral use, for example, as tablets, troches,lozenges, aqueous or oily suspensions, dispersible powders or granules,emulsions, hard or soft capsules, or syrups or elixirs. Compositionsintended for oral use may be prepared according to any method known tothe art for the manufacture of pharmaceutical compositions. Suchcompositions may contain one or more agents selected from sweeteningagents, flavoring agents, coloring agents and preserving agents in orderto provide pharmaceutically elegant and palatable preparations. Tabletscontain the active ingredient in admixture with other non-toxicpharmaceutically acceptable excipients which are suitable for themanufacture of tablets. These excipients may be, for example, inertdiluents, such as calcium carbonate, sodium carbonate, lactose, calciumphosphate or sodium phosphate; granulating and disintegrating agents,for example, corn starch, or alginic acid; binding agents, for examplestarch, gelatin or acacia, and lubricating agents, for example magnesiumstearate, stearic acid, or talc. The tablets may be uncoated or they maybe coated by known techniques to delay disintegration and absorption inthe gastrointestinal tract and thereby provide a sustained action over alonger period. For example, a time delay material such as glycerylmonostearate or glyceryl distearate may be employed. They may also becoated by the techniques described in U.S. Pat. Nos. 4,256,108,4,160,452, and 4,265,874 to form osmotic therapeutic tablets for controlrelease.

Compositions for oral use may also be presented as hard gelatin capsuleswherein the active ingredient is mixed with an inert solid diluent, forexample, calcium carbonate, calcium phosphate, or kaolin, or as softgelatin capsules wherein the active ingredient is mixed with water or anoil medium, for example peanut oil, liquid paraffin, or olive oil.

Aqueous suspensions contain the active materials in admixture withexcipients suitable for the manufacture of aqueous suspensions. Suchexcipients are suspending agents, for example sodiumcarboxymethylcellulose, methylcellulose, hydroxy-propylmethylcellulose,sodium alginate, polyvinyl-pyrrolidone, gum tragacanth and gum acacia;dispersing or wetting agents may be a naturally-occurring phosphatide,for example lecithin, or condensation products of an alkylene oxide withfatty acids, for example polyoxy-ethylene stearate, or condensationproducts of ethylene oxide with long chain aliphatic alcohols, forexample heptadecaethyleneoxycetanol, or condensation products ofethylene oxide with partial esters derived from fatty acids and ahexitol such as polyoxyethylene sorbitol monooleate, or condensationproducts of ethylene oxide with partial esters derived from fatty acidsand hexitol anhydrides, for example polyethylene sorbitan monooleate.The aqueous suspensions may also contain one or more preservatives, forexample ethyl, or n-propyl, p-hydroxybenzoate, one or more coloringagents, one or more flavoring agents, and one or more sweetening agents,such as sucrose or saccharin.

Oily suspensions may be formulated by suspending the active ingredientin a vegetable oil, for example arachis oil, olive oil, sesame oil, orcoconut oil, or in a mineral oil such as liquid paraffin. The oilysuspensions may contain a thickening agent, for example beeswax, hardparaffin, or cetyl alcohol. Sweetening agents such as those set forthabove, and flavoring agents may be added to provide a palatable oralpreparation. These compositions may be preserved by the addition of ananti-oxidant such as ascorbic acid.

Dispersible powders and granules suitable for preparation of an aqueoussuspension by the addition of water provide the active ingredient inadmixture with a dispersing or wetting agent, suspending agent and oneor more preservatives. Suitable dispersing or wetting agents andsuspending agents are exemplified by those already mentioned above.Additional excipients, for example sweetening, flavoring and coloringagents, may also be present.

The pharmaceutical compositions of the invention may also be in the formof oil-in-water emulsions. The oily phase may be a vegetable oil, forexample olive oil or arachis oil, or a mineral oil, for example liquidparaffin or mixtures of these. Suitable emulsifying agents may benaturally-occurring gums, for example gum acacia or gum tragacanth,naturally-occurring phosphatides, for example soy bean, lecithin, andesters or partial esters derived from fatty acids and hexitolanhydrides, for example sorbitan monooleate, and condensation productsof the said partial esters with ethylene oxide, for examplepolyoxyethylene sorbitan monooleate. The emulsions may also containsweetening and flavoring agents.

Syrups and elixirs may be formulated with sweetening agents, for exampleglycerol, propylene glycol, sorbitol or sucrose. Such compositions mayalso contain a demulcent, a preservative, and flavoring and coloringagents.

The pharmaceutical compositions may be in the form of a sterileinjectable aqueous or oleagenous suspension. This suspension may beformulated according to the known art using those suitable dispersing orwetting agents and suspending agents which have been mentioned above.The sterile injectable preparation may also be a sterile injectablesolution or suspension in a non-toxic parenterally acceptable diluent orsolvent, for example as a solution in 1,3-butane diol. Among theacceptable vehicles and solvents that may be employed are water,Ringer's solution, and isotonic sodium chloride solution. In addition,sterile, fixed oils are conventionally employed as a solvent orsuspending medium. For this purpose, any bland fixed oil may be employedincluding synthetic mono- or diglycerides. In addition, fatty acids suchas oleic acid find use in the preparation of injectables.

The pharmaceutical compositions may also be administered in the form ofsuppositories for rectal administration of the drug. These compositionscan be prepared by mixing the drug with a suitable non-irritatingexcipient which is solid at ordinary temperatures but liquid at therectal temperature and will therefore melt in the rectum to release thedrug. Such materials include, for example, cocoa butter and polyethyleneglycols.

For topical use, creams, ointments, jellies, solutions, or suspensions,etc., containing the compounds of the invention are employed. As usedherein, topical application is also meant to include the use ofmouthwashes and gargles.

The compounds of the invention may be used to treat or prevent variouskinase-related disorders. Thus, the present invention provides methodsfor treating or preventing such disorders. In some embodiments, theinvention provides a method for treating a kinase-mediated disorder in asubject that includes administering a therapeutically effective amountof a compound of any of the embodiments of the invention or apharmaceutical composition to the subject. In some embodiments, thesubject is a mammal, and in some such embodiments is a human. In someembodiments the disorder is mediated by CDK4, CDK6, and/or FLT3 complex.In some such embodiments, the disorder is mediated by CDK4 and/or CDK6.In other embodiments, the disease is mediated by FLT3. In someembodiments, administration of the compound, salt, or pharmaceuticalcomposition produces inhibition of CDK4 and FLT3. In some embodiments,the disorder is cancer. The present invention thus provides methods fortreating or preventing CDK4 and FLT3-mediated disease states, such ascancer. In some embodiments, the cancer is a tumor such as a solidtumor. However, in other embodiments, the cancer is a hematologicalcancer. In some embodiments, the cancer is acute myeloid leukemia. Insome embodiments, the disorder is mediated by p16 and p15 expression andin some such embodiments is correlated with low expression levels ofp15^(INK4B) and/or p16^(INK4A).

As described above, the compounds of the invention may also be used totreat proliferation-related disorders. Thus, the invention furtherprovides methods for treating such proliferation-related disorders in asubject. Such methods include administering to a subject in need thereofa therapeutically effective amount of the compound or pharmaceuticalcomposition of any of the embodiments. In some embodiments, the subjectis a mammal. In some such embodiments, the mammal is a human. In someembodiments, the proliferation-related disorder is abnormal cell growth.In other embodiments, the disorder is inflammation or aninflammation-related disorder. In still other embodiments, the disorderis a metabolic disease such as diabetes. In still other embodiments, thedisorder is cancer. In some such embodiments, the cancer is a solidtumor. In other such embodiments, the cancer is a hematological cancer,and, in some such embodiments, is acute myeloid leukemia.

The magnitude of a prophylactic or therapeutic dose of a compound of anyof the embodiments or a pharmaceutically acceptable salt, solvate,hydrate, or stereoisomer thereof in the acute or chronic treatment orprevention of a cancer or other disease or condition will vary with thenature and aggressiveness of the condition, and the route by which theactive ingredient is administered. The dose, and in some cases the dosefrequency, will also vary according to the condition to be treated, theage, body weight, and response of the individual patient. Suitabledosing regimens can be readily selected by those skilled in the art withdue consideration of such factors. In one embodiment, the doseadministered depends upon the specific compound to be used, and theweight and condition of the patient. In general, the dose per day is inthe range of from about 0.001 to 100 mg/kg, preferably about 1 to 25mg/kg, more preferably about 1 to about 5 mg/kg. For treatment of humanshaving a cancer, about 0.1 mg to about 15 g per day is administered inabout one to four divisions a day, preferably 10 mg to 12 g per day,more preferably from 40 mg to 500 mg per day. In one embodiment thecompounds of the invention are administered from 40 mg to 500 mg per dayin about one to four divisions a day. Additionally, the recommendeddaily dose can be administered in cycles as single agents or incombination with other therapeutic agents. In one embodiment, the dailydose is administered in a single dose or in equally divided doses. In arelated embodiment, the recommended daily dose can be administered onetime per week, two times per week, three times per week, four times perweek or five times per week. In some embodiments, a compound orpharmaceutical composition of the invention is dosed QD whereas in otherembodiments, it is dosed BID.

The compounds of the invention can be administered to provide systemicdistribution of the compound within the patient. Therefore, in someembodiments, the compounds of the invention are administered to producea systemic effect in the body.

The compounds of the invention may also be administered directly to asite affected by a condition, as, for example, an in the treatment of anaccessible area of skin or an esophageal cancer.

As indicated above, the compounds of the invention may be administeredvia oral, mucosal (including sublingual, buccal, rectal, nasal, orvaginal), parenteral (including subcutaneous, intramuscular, bolusinjection, intra-arterial, or intravenous), transdermal, or topicaladministration. In some embodiments, the compounds of the invention areadministered via mucosal (including sublingual, buccal, rectal, nasal,or vaginal), parenteral (including subcutaneous, intramuscular, bolusinjection, intra-arterial, or intravenous), transdermal, or topicaladministration. In other embodiments, the compounds of the invention areadministered via oral administration. In still other embodiments, thecompounds of the invention are not administered via oral administration.

Different therapeutically effective amounts may be applicable fordifferent conditions, as will be readily known by those of ordinaryskill in the art. Similarly, amounts sufficient to treat or prevent suchconditions, but insufficient to cause, or sufficient to reduce, adverseeffects associated with conventional therapies are also encompassed bythe above described dosage amounts and dose frequency schedules.

Some methods of the invention comprise the administration of a compoundof the invention and an additional therapeutic agent (i.e., atherapeutic agent other than a compound of the invention). Thus, thecompounds of the invention can be used in combination with at least oneother therapeutic agent, for example with a second therapeutic agent.Examples of additional therapeutic agents include, but are not limitedto, antibiotics, anti-emetic agents, antidepressants, antifungal agents,anti-inflammatory agents, antineoplastic agents, antiviral agents,cytotoxic agents, and other anticancer agents, immunomodulatory agents,alpha-interferons, β-interferons, alkylating agents, hormones, andcytokines. In one embodiment, the invention encompasses administrationof an additional therapeutic agent that demonstrates anti-canceractivity. In another embodiment, an additional therapeutic agent thatdemonstrates cytotoxic activity is administered to a subject such as acancer patient. In some embodiments, the second therapeutic agent isselected from cytosine arabinoside, daunorubicin, idarubicin,doxorubicin, cyclophosphamide, etoposide, carboplatin, fludarabine,mitoxantrone, dexamethasone, rituximab, midostaurin, a granulocytecolony-stimulating factor, filgrastim, PEG-filgrastim, lenograstim,decitabine, azacitidine, paclitaxel, gemcitibine, motesanibdisphosphate, panitumumab, an antibody directed against CD33, or a CD33bispecific T-cell engager antibody. In other embodiments, the secondtherapeutic agent is selected from cytosine arabinoside, daunorubicin,idarubicin, doxorubicin, cyclophosphamide, etoposide, carboplatin,fludarabine, mitoxantrone, dexamethasone, rituximab, midostaurin, agranulocyte colony-stimulating factor, filgrastim, PEG-filgrastim,lenograstim, decitabine, azacitidine, paclitaxel, gemcitibine, motesanibdisphosphate, panitumumab. In still other such embodiments, the secondtherapeutic agent is selected from cytosine arabinoside, daunorubicin,idarubicin, doxorubicin, cyclophosphamide, etoposide, carboplatin,fludarabine, mitoxantrone, dexamethasone, rituximab, midostaurin,decitabine, azacitidine, paclitaxel, gemcitibine, or motesanibdisphosphate. In some such embodiments, the second therapeutic agent iscytosine arabinoside. In other embodiments, the second therapeutic agentis daunorubicin, idarubicin, or doxorubicin. In still other embodiments,the second therapeutic agent is azacitidine or decitabine.

The compounds of the invention and the other therapeutics agent can actadditively or, preferably, synergistically. In some embodiments, acomposition comprising a compound of the invention is administeredconcurrently with the administration of another therapeutic agent, whichcan be part of the same composition or can be in a different compositionfrom the one that comprises the compound of the invention. In otherembodiments, a compound of the invention is administered prior to, orsubsequent to, administration of another therapeutic agent. In stillother embodiments, a compound of the invention is administered to apatient who has not previously undergone or is not currently undergoingtreatment with another therapeutic agent. A compound of the inventionmay be administered to a subject that has had, is currently undergoing,or is scheduled to receive radiation therapy. In some such embodiments,the subject is a cancer patient.

When administered as a combination, the therapeutic agents can beformulated as separate compositions that are administered at the sametime or sequentially at different times, or the therapeutic agents canbe given as a single composition. The phrase “co-therapy” (or“combination-therapy”), in defining use of a compound of the presentinvention and another pharmaceutical agent, is intended to embraceadministration of each agent in a sequential manner in a regimen thatwill provide beneficial effects of the drug combination, and is intendedas well to embrace co-administration of these agents in a substantiallysimultaneous manner, such as in a single capsule having a fixed ratio ofthese active agents or in multiple, separate capsules for each agent.Specifically, the administration of compounds of the present inventionmay be in conjunction with additional therapies known to those skilledin the art in the prevention or treatment of neoplasia, such as withradiation therapy or with cytostatic or cytotoxic agents.

If formulated as a fixed dose, such combination products employ thecompounds of this invention within the accepted dosage ranges. Compoundsof any of the embodiments described herein may also be administeredsequentially with known anticancer or cytotoxic agents when acombination formulation is inappropriate. The invention is not limitedin the sequence of administration as compounds of the invention may beadministered either prior to, simultaneous with, or after administrationof a known anticancer or cytotoxic agent.

There are large numbers of antineoplastic agents available in commercialuse, in clinical evaluation and in pre-clinical development, which maybe selected for treatment of neoplasia by combination drug chemotherapy.Such antineoplastic agents fall into several major categories, namely,antibiotic-type agents, alkylating agents, antimetabolite agents,hormonal agents, immunological agents, interferon-type agents and acategory of miscellaneous agents.

A first family of antineoplastic agents which may be used in combinationwith compounds of the present invention consists ofantimetabolite-type/thymidilate synthase inhibitor antineoplasticagents. Suitable antimetabolite antineoplastic agents may be selectedfrom, but are not limited to, the group consisting of 5-FU-fibrinogen,acanthifolic acid, aminothiadiazole, brequinar sodium, carmofur,Ciba-Geigy CGP-30694, cyclopentyl cytosine, cytarabine phosphatestearate, cytarabine conjugates, Lilly DATHF, Merrel Dow DDFC,dezaguanine, dideoxycytidine, dideoxyguanosine, didox, Yoshitomi DMDC,doxifluridine, Wellcome EHNA, Merck & Co. EX-015, fazarabine,floxuridine, fludarabine phosphate, 5-fluorouracil,N-(2′-furanidyl)-5-fluorouracil, Daiichi Seiyaku FO-152, isopropylpyrrolizine, Lilly LY-188011, Lilly LY-264618, methobenzaprim,methotrexate, Wellcome MZPES, norspermidine, NCI NSC-127716, NCINSC-264880, NCI NSC-39661, NCI NSC-612567, Warner-Lambert PALA,pentostatin, piritrexim, plicamycin, Asahi Chemical PL-AC, TakedaTAC-788, thioguanine, tiazofurin, Erbamont TIF, trimetrexate, tyrosinekinase inhibitors, Taiho UFT, and uricytin.

A second family of antineoplastic agents which may be used incombination with compounds of the present invention consists ofalkylating-type antineoplastic agents. Suitable alkylating-typeantineoplastic agents may be selected from, but are not limited to, thegroup consisting of Shionogi 254-S, aldo-phosphamide analogues,altretamine, anaxirone, Boehringer Mannheim BBR-2207, bestrabucil,budotitane, Wakunaga CA-102, carboplatin, carmustine, Chinoin-139,Chinoin-153, chlorambucil, cisplatin, cyclophosphamide, AmericanCyanamid CL-286558, Sanofi CY-233, cyplatate, Degussa D-19-384, SumimotoDACHP(Myr)₂, diphenylspiromustine, diplatinum cytostatic, Erbadistamycin derivatives, Chugai DWA-2114R, ITI E09, elmustine, ErbamontFCE-24517, estramustine phosphate sodium, fotemustine, Unimed G-6-M,Chinoin GYKI-17230, hepsul-fam, ifosfamide, iproplatin, lomustine,mafosfamide, mitolactol, Nippon Kayaku NK-121, NCI NSC-264395, NCINSC-342215, oxaliplatin, Upjohn PCNU, prednimustine, Proter PTT-119,ranimustine, semustine, SmithKline SK&F-101772, Yakult Honsha SN-22,spiromus-tine, Tanabe Seiyaku TA-077, tauromustine, temozolomide,teroxirone, tetraplatin, and trimelamol.

A third family of antineoplastic agents which may be used in combinationwith compounds of the present invention consists of antibiotic-typeantineoplastic agents. Suitable antibiotic-type antineoplastic agentsmay be selected from, but are not limited to, the group consisting ofTaiho 4181-A, aclarubicin, actinomycin D, actinoplanone, ErbamontADR-456, aeroplysinin derivative, Ajinomoto AN-201-II, Ajinomoto AN-3,Nippon Soda anisomycins, anthracycline, azino-mycin-A, bisucaberin,Bristol-Myers BL-6859, Bristol-Myers BMY-25067, Bristol-Myers BMY-25551,Bristol-Myers BMY-26605, Bristol-Myers BMY-27557, Bristol-MyersBMY-28438, bleomycin sulfate, bryostatin-1, Taiho C-1027, calichemycin,chromoximycin, dactinomycin, daunorubicin, Kyowa Hakko DC-102, KyowaHakko DC-79, Kyowa Hakko DC-88A, Kyowa Hakko DC89-A1, Kyowa HakkoDC92-B, ditrisarubicin B, Shionogi DOB-41, doxorubicin,doxorubicin-fibrinogen, elsamicin-A, epirubicin, erbstatin, esorubicin,esperamicin-Al, esperamicin-Alb, Erbamont FCE-21954, Fujisawa FK-973,fostriecin, Fujisawa FR-900482, glidobactin, gregatin-A, grincamycin,herbimycin, idarubicin, illudins, kazusamycin, kesarirhodins, KyowaHakko KM-5539, Kirin Brewery KRN-8602, Kyowa Hakko KT-5432, Kyowa HakkoKT-5594, Kyowa Hakko KT-6149, American Cyanamid LL-D49194, Meiji SeikaME 2303, menogaril, mitomycin, mitoxantrone, SmithKline M-TAG,neoenactin, Nippon Kayaku NK-313, Nippon Kayaku NKT-01, SRIInternational NSC-357704, oxalysine, oxaunomycin, peplomycin, pilatin,pirarubicin, porothramycin, pyrindanycin A, Tobishi RA-I, rapamycin,rhizoxin, rodorubicin, sibanomicin, siwenmycin, Sumitomo SM-5887, SnowBrand SN-706, Snow Brand SN-07, sorangicin-A, sparsomycin, SSPharmaceutical SS-21020, SS Pharmaceutical SS-7313B, SS PharmaceuticalSS-9816B, steffimycin B, Taiho 4181-2, talisomycin, Takeda TAN-868A,terpentecin, thrazine, tricrozarin A, Upjohn U-73975, Kyowa HakkoUCN-10028A, Fujisawa WF-3405, Yoshitomi Y-25024, and zorubicin.

A fourth family of antineoplastic agents which may be used incombination with compounds of the present invention consists of amiscellaneous family of antineoplastic agents, including tubulininteracting agents, topoisomerase II inhibitors, topoisomerase Iinhibitors and hormonal agents, selected from, but not limited to, thegroup consisting of α-carotene, α-difluoromethyl-arginine, acitretin,Biotec AD-5, Kyorin AHC-52, alstonine, amonafide, amphethinile,amsacrine, Angiostat, ankinomycin, anti-neoplaston A10, antineoplastonA2, antineoplaston A3, antineoplaston A5, antineoplaston AS2-1, HenkelAPD, aphidicolin glycinate, asparaginase, Avarol, baccharin, batracylin,benfluoron, benzotript, Ipsen-Beaufour BIM-23015, bisantrene,Bristol-Myers BMY-40481, Vestar boron-10, bromofosfamide, WellcomeBW-502, Wellcome BW-773, caracemide, carmethizole hydrochloride,Ajinomoto CDAF, chlorsulfaquinoxalone, Chemex CHX-2053, Chemex CHX-100,Warner-Lambert CI-921, Warner-Lambert CI-937, Warner-Lambert CI-941,Warner-Lambert CI-958, clanfenur, claviridenone, ICN compound 1259, ICNcompound 4711, Contracan, Yakult Honsha CPT-11, crisnatol, curaderm,cytochalasin B, cytarabine, cytocytin, Merz D-609, DABIS maleate,dacarbazine, datelliptinium, didemnin-B, dihaematoporphyrin ether,dihydrolenperone, dinaline, distamycin, Toyo Pharmar DM-341, ToyoPharmar DM-75, Daiichi Seiyaku DN-9693, docetaxel elliprabin,elliptinium acetate, Tsumura EPMTC, the epothilones, ergotamine,etoposide, etretinate, fenretinide, Fujisawa FR-57704, gallium nitrate,genkwadaphnin, Chugai GLA-43, Glaxo GR-63178, grifolan NMF-5N,hexadecylphosphocholine, Green Cross HO-221, homoharringtonine,hydroxyurea, BTG ICRF-187, ilmofosine, isoglutamine, isotretinoin,Otsuka JI-36, Ramot K-477, Otsuak K-76COONa, Kureha Chemical K-AM, MECTCorp KI-8110, American Cyanamid L-623, leukoregulin, lonidamine,Lundbeck LU-23-112, Lilly LY-186641, NCl (US) MAP, marycin, Merrel DowMDL-27048, Medco MEDR-340, merbarone, merocyanlne derivatives,methylanilinoacridine, Molecular Genetics MGI-136, minactivin,mitonafide, mitoquidone mopidamol, motretinide, Zenyaku Kogyo MST-16,N-(retinoyl)amino acids, Nisshin Flour Milling N-021,N-acylated-dehydroalanines, nafazatrom, Taisho NCU-190, nocodazolederivative, Normosang, NCI NSC-145813, NCI NSC-361456, NCI NSC-604782,NCI NSC-95580, ocreotide, Ono ONO-112, oquizanocine, Akzo Org-10172,paclitaxel, pancratistatin, pazelliptine, Warner-Lambert PD-111707,Warner-Lambert PD-115934, Warner-Lambert PD-131141, Pierre FabrePE-1001, ICRT peptide D, piroxantrone, polyhaematoporphyrin, polypreicacid, Efamol porphyrin, probimane, procarbazine, proglumide, Invitronprotease nexin I, Tobishi RA-700, razoxane, Sapporo Breweries RBS,restrictin-P, retelliptine, retinoic acid, Rhone-Poulenc RP-49532,Rhone-Poulenc RP-56976, SmithKline SK&F-104864, Sumitomo SM-108, KuraraySMANCS, SeaPharm SP-10094, spatol, spirocyclopropane derivatives,spirogermanium, Unimed, SS Pharmaceutical SS-554, strypoldinone,Stypoldione, Suntory SUN 0237, Suntory SUN 2071, superoxide dismutase,Toyama T-506, Toyama T-680, taxol, Teijin TEI-0303, teniposide,thaliblastine, Eastman Kodak TJB-29, tocotrienol, topotecan, Topostin,Teijin TT-82, Kyowa Hakko UCN-01, Kyowa Hakko UCN-1028, ukrain, EastmanKodak USB-006, vinblastine sulfate, vincristine, vindesine,vinestramide, vinorelbine, vintriptol, vinzolidine, withanolides, andYamanouchi YM-534.

Alternatively, the present compounds may also be used in co-therapieswith other anti-neoplastic agents, such as acemannan, aclarubicin,aldesleukin, alemtuzumab, alitretinoin, altretamine, amifostine,aminolevulinic acid, amrubicin, amsacrine, anagrelide, anastrozole,ANCER, ancestim, ARGLABIN, arsenic trioxide, BAM 002 (Novelos),bexarotene, bicalutamide, broxuridine, capecitabine, celmoleukin,cetrorelix, cladribine, clotrimazole, cytarabine ocfosfate, DA 3030(Dong-A), daclizumab, denileukin diftitox, deslorelin, dexrazoxane,dilazep, docetaxel, docosanol, doxercalciferol, doxifluridine,doxorubicin, bromocriptine, carmustine, cytarabine, fluorouracil, HITdiclofenac, interferon alfa, daunorubicin, doxorubicin, tretinoin,edelfosine, edrecolomab, eflornithine, emitefur, epirubicin, epoetinbeta, etoposide phosphate, exemestane, exisulind, fadrozole, filgrastim,finasteride, fludarabine phosphate, formestane, fotemustine, galliumnitrate, gemcitabine, gemtuzumab zogamicin, gimeracil/oteracil/tegafurcombination, glycopine, goserelin, heptaplatin, human chorionicgonadotropin, human fetal alpha fetoprotein, ibandronic acid,idarubicin, (imiquimod, interferon alfa, interferon alfa, natural,interferon alfa-2, interferon alfa-2a, interferon alfa-2b, interferonalfa-N1, interferon alfa-n3, interferon alfacon-1, interferon alpha,natural, interferon beta, interferon beta-1a, interferon beta-1b,interferon gamma, natural interferon gamma-1a, interferon gamma-1b,interleukin-1 beta, iobenguane, irinotecan, irsogladine, lanreotide, LC9018 (Yakult), leflunomide, lenograstim, lentinan sulfate, letrozole,leukocyte alpha interferon, leuprorelin, levamisole+fluorouracil,liarozole, lobaplatin, lonidamine, lovastatin, masoprocol, melarsoprol,metoclopramide, mifepristone, miltefosine, mirimostim, mismatched doublestranded RNA, mitoguazone, mitolactol, mitoxantrone, molgramostim,nafarelin, naloxone+pentazocine, nartograstim, nedaplatin, nilutamide,noscapine, novel erythropoiesis stimulating protein, NSC 631570octreotide, oprelvekin, osaterone, oxaliplatin, paclitaxel, pamidronicacid, pegaspargase, peginterferon alfa-2b, pentosan polysulfate sodium,pentostatin, picibanil, pirarubicin, rabbit antithymocyte polyclonalantibody, polyethylene glycol interferon alfa-2a, porfimer sodium,raloxifene, raltitrexed, rasburicase, rhenium Re 186 etidronate, RIIretinamide, rituximab, romurtide, samarium (153 Sm) lexidronam,sargramostim, sizofuran, sobuzoxane, sonermin, strontium-89 chloride,suramin, tasonermin, tazarotene, tegafur, temoporfin, temozolomide,teniposide, tetrachlorodecaoxide, thalidomide, thymalfasin, thyrotropinalfa, topotecan, toremifene, tositumomab-iodine 131, trastuzumab,treosulfan, tretinoin, trilostane, trimetrexate, triptorelin, tumornecrosis factor alpha, natural, ubenimex, bladder cancer vaccine,Maruyama vaccine, melanoma lysate vaccine, valrubicin, verteporfin,vinorelbine, VIRULIZIN, zinostatin stimalamer, or zoledronic acid;abarelix; AE 941 (Aeterna), ambamustine, antisense oligonucleotide,bc1-2 (Genta), APC 8015 (Dendreon), cetuximab, decitabine,dexaminoglutethimide, diaziquone, EL 532 (Elan), EM 800 (Endorecherche),eniluracil, etanidazole, fenretinide, filgrastim SDO1 (Amgen),fulvestrant, galocitabine, gastrin 17 immunogen, HLA-B7 gene therapy(Vical), granulocyte macrophage colony stimulating factor, histaminedihydrochloride, ibritumomab tiuxetan, ilomastat, IM 862 (Cytran),interleukin-2, iproxifene, LDI 200 (Milkhaus), leridistim, lintuzumab,CA 125 MAb (Biomira), cancer MAb (Japan Pharmaceutical Development),HER-2 and Fc MAb (Medarex), idiotypic 105AD7 MAb (CRC Technology),idiotypic CEA MAb (Trilex), LYM-1-iodine 131 MAb (Techniclone),polymorphic epithelial mucin-yttrium 90 MAb (Antisoma), marimastat,menogaril, mitumomab, motexafin gadolinium, MX 6 (Galderma), nelarabine,nolatrexed, P 30 protein, pegvisomant, pemetrexed, porfiromycin,prinomastat, RL 0903 (Shire), rubitecan, satraplatin, sodiumphenylacetate, sparfosic acid, SRL 172 (SR Pharma), SU 5416 (SUGEN), TA077 (Tanabe), tetrathiomolybdate, thaliblastine, thrombopoietin, tinethyl etiopurpurin, tirapazamine, cancer vaccine (Biomira), melanomavaccine (New York University), melanoma vaccine (Sloan KetteringInstitute), melanoma oncolysate vaccine (New York Medical College),viral melanoma cell lysates vaccine (Royal Newcastle Hospital), orvalspodar.

The compounds of the invention may further be used with VEGFRinhibitors. Other compounds described in the following patents andpatent applications can be used in combination therapy: U.S. Pat. No.6,258,812, US 2003/0105091, WO 01/37820, U.S. Pat. No. 6,235,764, WO01/32651, U.S. Pat. No. 6,630,500, U.S. Pat. No. 6,515,004, U.S. Pat.No. 6,713,485, U.S. Pat. No. 5,521,184, U.S. Pat. No. 5,770,599, U.S.Pat. No. 5,747,498, WO 02/68406, WO 02/66470, WO 02/55501, WO 04/05279,WO 04/07481, WO 04/07458, WO 04/09784, WO 02/59110, WO 99/45009, WO00/59509, WO 99/61422, U.S. Pat. No. 5,990,141, WO 00/12089, and WO00/02871.

In some embodiments, the combination comprises a composition of thepresent invention in combination with at least one anti-angiogenicagent. Agents are inclusive of, but not limited to, in vitrosynthetically prepared chemical compositions, antibodies, antigenbinding regions, radionuclides, and combinations and conjugates thereof.An agent can be an agonist, antagonist, allosteric modulator, toxin or,more generally, may act to inhibit or stimulate its target (e.g.,receptor or enzyme activation or inhibition), and thereby promote celldeath or arrest cell growth.

Exemplary anti-tumor agents include HERCEPTIN™ (trastuzumab), which maybe used to treat breast cancer and other forms of cancer, and RITUXAN™(rituximab), ZEVALIN™ (ibritumomab tiuxetan), and LYMPHOCIDE™(epratuzumab), which may be used to treat non-Hodgkin's lymphoma andother forms of cancer, GLEEVAC™ which may be used to treat chronicmyeloid leukemia and gastrointestinal stromal tumors, and BEXXAR™(iodine 131 tositumomab) which may be used for treatment ofnon-Hodgkins's lymphoma.

Exemplary anti-angiogenic agents include ERBITUX™ (IMC-C225), KDR(kinase domain receptor) inhibitory agents (e.g., antibodies and antigenbinding regions that specifically bind to the kinase domain receptor),anti-VEGF agents (e.g., antibodies or antigen binding regions thatspecifically bind VEGF, or soluble VEGF receptors or a ligand bindingregion thereof) such as AVASTIN™ or VEGF-TRAP™, and anti-VEGF receptoragents (e.g., antibodies or antigen binding regions that specificallybind thereto), EGFR inhibitory agents (e.g., antibodies or antigenbinding regions that specifically bind thereto) such as ABX-EGF(panitumumab), IRESSA™ (gefitinib), TARCEVA™ (erlotinib), anti-Ang 1 andanti-Ang2 agents (e.g., antibodies or antigen binding regionsspecifically binding thereto or to their receptors, e.g., Tie2/Tek), andanti-Tie2 kinase inhibitory agents (e.g., antibodies or antigen bindingregions that specifically bind thereto). The pharmaceutical compositionsof the present invention can also include one or more agents (e.g.,antibodies, antigen binding regions, or soluble receptors) thatspecifically bind and inhibit the activity of growth factors, such asantagonists of hepatocyte growth factor (HGF, also known as ScatterFactor), and antibodies or antigen binding regions that specificallybind its receptor “c-met”.

Other anti-angiogenic agents include Campath, IL-8, B-FGF, Tekantagonists (Ceretti et al., U.S. Publication No. 2003/0162712; U.S.Pat. No. 6,413,932), anti-TWEAK agents (e.g., specifically bindingantibodies or antigen binding regions, or soluble TWEAK receptorantagonists; see, Wiley, U.S. Pat. No. 6,727,225), ADAM distintegrindomain to antagonize the binding of integrin to its ligands (Fanslow etal., U.S. Publication No. 2002/0042368), specifically binding anti-ephreceptor and/or anti-ephrin antibodies or antigen binding regions (U.S.Pat. Nos. 5,981,245; 5,728,813; 5,969,110; 6,596,852; 6,232,447;6,057,124 and patent family members thereof), and anti-PDGF-BBantagonists (e.g., specifically binding antibodies or antigen bindingregions) as well as antibodies or antigen binding regions specificallybinding to PDGF-BB ligands, and PDGFR kinase inhibitory agents (e.g.,antibodies or antigen binding regions that specifically bind thereto).

Additional anti-angiogenic/anti-tumor agents include: SD-7784 (Pfizer,USA); cilengitide. (Merck KGaA, Germany, EPO 770622); pegaptaniboctasodium, (Gilead Sciences, USA); Alphastatin, (BioActa, UK); M-PGA,(Celgene, USA, U.S. Pat. No. 5,712,291); ilomastat, (Arriva, USA, U.S.Pat. No. 5,892,112); emaxanib, (Pfizer, USA, U.S. Pat. No. 5,792,783);vatalanib, (Novartis, Switzerland); 2-methoxyestradiol, (EntreMed, USA);TLC ELL-12, (Elan, Ireland); anecortave acetate, (Alcon, USA); alpha-D148 Mab, (Amgen, USA); CEP-7055, (Cephalon, USA); anti-Vn Mab, (Crucell,Netherlands) DAC: antiangiogenic, (ConjuChem, Canada); Angiocidin,(InKine Pharmaceutical, USA); KM-2550, (Kyowa Hakko, Japan); SU-0879,(Pfizer, USA); CGP-79787, (Novartis, Switzerland, EP 970070); ARGENTtechnology, (Ariad, USA); YIGSR-Stealth, (Johnson & Johnson, USA);fibrinogen-E fragment, (BioActa, UK); angiogenesis inhibitor, (Trigen,UK); TBC-1635, (Encysive Pharmaceuticals, USA); SC-236, (Pfizer, USA);ABT-567, (Abbott, USA); Metastatin, (EntreMed, USA); angiogenesisinhibitor, (Tripep, Sweden); maspin, (Sosei, Japan); 2-methoxyestradiol,(Oncology Sciences Corporation, USA); ER-68203-00, (IVAX, USA); Benefin,(Lane Labs, USA); Tz-93, (Tsumura, Japan); TAN-1120, (Takeda, Japan);FR-111142, (Fujisawa, Japan, JP 02233610); platelet factor 4, (RepliGen,USA, EP 407122); vascular endothelial growth factor antagonist, (Borean,Denmark); cancer therapy, (University of South Carolina, USA);bevacizumab (pINN), (Genentech, USA); angiogenesis inhibitors, (SUGEN,USA); XL 784, (Exelixis, USA); XL 647, (Exelixis, USA); MAb, alpha5beta3integrin, second generation, (Applied Molecular Evolution, USA andMedImmune, USA); gene therapy, retinopathy, (Oxford BioMedica, UK);enzastaurin hydrochloride (USAN), (Lilly, USA); CEP 7055, (Cephalon, USAand Sanofi-Synthelabo, France); BC 1, (Genoa Institute of CancerResearch, Italy); angiogenesis inhibitor, (Alchemia, Australia); VEGFantagonist, (Regeneron, USA); rBPI 21 and BPI-derived antiangiogenic,(XOMA, USA); PI 88, (Progen, Australia); cilengitide (pINN), (MerckKGaA, German; Munich Technical University, Germany, Scripps Clinic andResearch Foundation, USA); cetuximab (INN), (Aventis, France); AVE 8062,(Ajinomoto, Japan); AS1404, (Cancer Research Laboratory, New Zealand);SG 292, (Telios, USA); Endostatin, (Boston Childrens Hospital, USA); ATN161, (Attenuon, USA); ANGIOSTATIN, (Boston Childrens Hospital, USA);2-methoxyestradiol, (Boston Childrens Hospital, USA); ZD 6474,(AstraZeneca, UK); ZD 6126, (Angiogene Pharmaceuticals, UK); PPI 2458,(Praecis, USA); AZD 9935, (AstraZeneca, UK); AZD 2171, (AstraZeneca,UK); vatalanib (pINN), (Novartis, Switzerland and Schering AG, Germany);tissue factor pathway inhibitors, (EntreMed, USA); pegaptanib (Pinn),(Gilead Sciences, USA); xanthorrhizol, (Yonsei University, South Korea);vaccine, gene-based, VEGF-2, (Scripps Clinic and Research Foundation,USA); SPV5.2, (Supratek, Canada); SDX 103, (University of California atSan Diego, USA); PX 478, (ProlX, USA); METASTATIN, (EntreMed, USA);troponin I, (Harvard University, USA); SU 6668, (SUGEN, USA); OXI 4503,(OXiGENE, USA); o-guanidines, (Dimensional Pharmaceuticals, USA);motuporamine C, (British Columbia University, Canada); CDP 791,(Celltech Group, UK); atiprimod (pINN), (GlaxoSmithKline, UK); E 7820,(Eisai, Japan); CYC 381, (Harvard University, USA); AE 941, (Aeterna,Canada); vaccine, angiogenesis, (EntreMed, USA); urokinase plasminogenactivator inhibitor, (Dendreon, USA); oglufanide (pINN), (Melmotte,USA); HIF-1alfa inhibitors, (Xenova, UK); CEP 5214, (Cephalon, USA); BAYRES 2622, (Bayer, Germany); Angiocidin, (InKine, USA); A6, (Angstrom,USA); KR 31372, (Korea Research Institute of Chemical Technology, SouthKorea); GW 2286, (GlaxoSmithKline, UK); EHT 0101, (ExonHit, France); CP868596, (Pfizer, USA); CP 564959, (OSI, USA); CP 547632, (Pfizer, USA);786034, (GlaxoSmithKline, UK); KRN 633, (Kirin Brewery, Japan); drugdelivery system, intraocular, 2-methoxyestradiol, (EntreMed, USA);anginex, (Maastricht University, Netherlands, and Minnesota University,USA); ABT 510, (Abbott, USA); AAL 993, (Novartis, Switzerland); VEGI,(ProteomTech, USA); tumor necrosis factor-alpha inhibitors, (NationalInstitute on Aging, USA); SU 11248, (Pfizer, USA and SUGEN USA); ABT518, (Abbott, USA); YH16, (Yantai Rongchang, China); S-3APG, (BostonChildrens Hospital, USA and EntreMed, USA); MAb, KDR, (ImClone Systems,USA); MAb, alpha5 beta1, (Protein Design, USA); KDR kinase inhibitor,(Celltech Group, UK, and Johnson & Johnson, USA); GFB 116, (SouthFlorida University, USA and Yale University, USA); CS 706, (Sankyo,Japan); combretastatin A4 prodrug, (Arizona State University, USA);chondroitinase AC, (IBEX, Canada); BAY RES 2690, (Bayer, Germany); AGM1470, (Harvard University, USA, Takeda, Japan, and TAP, USA); AG 13925,(Agouron, USA); Tetrathiomolybdate, (University of Michigan, USA);GCS100, (Wayne State University, USA) CV 247, (Ivy Medical, UK); CKD732, (Chong Kun Dang, South Korea); MAb, vascular endothelium growthfactor, (Xenova, UK); irsogladine (INN), (Nippon Shinyaku, Japan); RG13577, (Aventis, France); WX 360, (Wilex, Germany); squalamine (pINN),(Genaera, USA); RPI 4610, (Sirna, USA); cancer therapy, (Marinova,Australia); heparanase inhibitors, (InSight, Israel); KL 3106, (Kolon,South Korea); Honokiol, (Emory University, USA); ZK CDK, (Schering AG,Germany); ZK Angio, (Schering AG, Germany); ZK 229561, (Novartis,Switzerland, and Schering AG, Germany); XMP 300, (XOMA, USA); VGA 1102,(Taisho, Japan); VEGF receptor modulators, (Pharmacopeia, USA);VE-cadherin-2 antagonists, (ImClone Systems, USA); Vasostatin, (NationalInstitutes of Health, USA); vaccine, Flk-1, (ImClone Systems, USA); TZ93, (Tsumura, Japan); TumStatin, (Beth Israel Hospital, USA); truncatedsoluble FLT1 (vascular endothelial growth factor receptor 1), (Merck &Co, USA); Tie-2 ligands, (Regeneron, USA); and, thrombospondin 1inhibitor, (Allegheny Health, Education and Research Foundation, USA).

Alternatively, the present compounds may also be used in co-therapieswith other anti-neoplastic agents, such as VEGF antagonists, otherkinase inhibitors including p38 inhibitors, aurora kinase inhibitors,c-met inhibitors, KDR inhibitors, EGF inhibitors and CDK inhibitors, TNFinhibitors, matrix metalloproteinases (MMP) inhibitors, COX-2 inhibitorsincluding celecoxib, NSAID's, or α_(v)β₃ inhibitors.

The compounds of the invention can be prepared using the generalsynthetic routes shown in the following schemes and described more fullyin the Examples.

Scheme 1 illustrates how compounds of the invention can be preparedstarting from chloropyrimidine 1A. Reaction of chloropyrimidine 1A withan appropriate amine provides diaminopyridimine 1B which can beiodinated to form 1C with N-iodosuccinimide. Iodopyrimidine 1C can bereacted with the appropriate zinc reagent 1D to form the unfusedbicyclic intermediate 1E which can then be cyclized to form the usefulreagent 1F. Reaction of 1F with suitable chloro substituted andprotected R¹ groups or precursors (See Scheme 3) may be used to convert1F to 1G which may then be converted into various compounds of theinvention as shown in Scheme 4.

Scheme 2 provides an alternative route showing how compounds of theinvention can be prepared starting from bromodichloropyrimidine 2A.Reaction of bromodichloropyrimidine 2A with an appropriate amineprovides aminobromochloropyridimine 2B which can be converted todiaminobromopyrimidine 2C by reaction with ammonium hydroxide inisopropanol. Diaminobromopyrimidine 2C can be reacted with the boronicacid reagent 2D to form the unfused bicyclic intermediate 2E which canthen be cyclized to form the useful reagent 2F. Reaction of 2F withsuitable chloro substituted and protected R¹ groups or precursors (SeeScheme 3) may be used to convert 2F to 2G which may then be convertedinto various compounds of the invention as shown in Scheme 4.

Scheme 3 shows how various R^(1′)—Cl Boc protected compounds may beprepared from the starting amine The Boc-protected chloro-substitutedcompounds 3B, 3D, 3F, and 3H are useful reagents for preparing compoundsof the invention as shown in Schemes 1 and 2.

Scheme 4 shows how Boc-protected compound 4A prepared from Boc-protectedchloro compound 3B and 1F (see Scheme 1) can be deprotected to form 4B.The deprotected 4B may be used to prepare a wide variety of derivativesas shown in Scheme 4. For example, reaction of 4B with acetic anhydridecan be used to prepare 4C. Similarly 4B can be reacted with commerciallyavailable (t-butyldimethylsilyloxy)acetaldehyde and(t-butyldimethylsilyloxy)propanal and then be reduced to respectivelyform 4D and 4E. As described in Example 5, compounds such as 4B may bereacted with 2,5-dioxopyrrolidin-1-yl2-acetoxyacetate to form 4F aftertreatment with sodium methoxide. 4B may also be used to form compoundssuch as 4G by treatment with methylchlorooxoacetate followed by reactionwith LiOH in a mixture of tetrahydrofuran, methanol, and water. As afinal, but non-exhaustive example, compounds such as 4B may be treatedwith reagents such as commercially available 3,3,3-trifluoropropanal andthen reduced to form trifluoromethyl compounds such as 4H. Various othercompounds can be prepared from compound 4B. For example, a reagent suchas an alkyl chloroformate, for example ethyl chloroformate, may bereacted with the deprotected compound 4B to produce compounds in whichR⁶ is —C(═O)—O—(C₁-C₆ alkyl).

The invention is further described by reference to the followingexamples, which are intended to exemplify the claimed invention but notto limit it in any way.

EXAMPLES

Unless otherwise stated, all starting materials were obtained fromcommercial sources such as Sigma-Aldrich, St. Louis, Mo., or wereobtained using literature procedures.

¹H-NMR spectra were typically acquired on a Bruker Avance III 500spectrometer system (Bruker, Bilerica, Mass.) operating at a ¹Hfrequency of 500.13 MHz, equipped with a Bruker 5 mm PABBI probe with az-axis gradient; or on a Bruker Avance II 400 spectrometer operating ata ¹H frequency of 400.23 MHz, equipped with a Bruker 5 mm PABBO probewith a z-axis gradient. Samples were typically dissolved in 500 μLDMSO-d₆, CD₃OD, CDCl₃, or another deuterated NMR solvent for NMRanalysis. ¹H chemical shifts are referenced to the residual solventsignals from DMSO-d₆ at δ 2.50, CD₃OD at δ 3.30, or other referencesolvents, or may be referenced to tetramethylsilane. Significant peakswere tabulated and typically include: number of protons, multiplicity(s, singlet; d, doublet; dd, doublet of doublets; t, triplet; q,quartet; m, multiplet; br s, broad singlet) and coupling constant(s) inHertz.

Electron Ionization (EI) mass spectra were typically recorded on anAgilent Technologies 6140 Quadrupole LC/MS mass spectrometer. Massspectrometry results are reported as the ratio of mass over charge,sometimes followed by the relative abundance of each ion (inparentheses).

The following Abbreviations are used to refer to various reagents andsolvents:

-   -   Ac₂O Acetic anhydride    -   AcOH Acetic acid    -   ATP Adenosine Triphosphate    -   BSA Bovine Serum Albumin    -   n-BuLi n-butyllithium    -   DCM Dichloromethane    -   DMEM Dulbecco's Modified Eagle Medium    -   DMF N,N-Dimethylformamide    -   DMSO Dimethylsulfoxide    -   DTT Dithiothreitol    -   EDC N-(3-dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride    -   EDTA Ethyelenediaminetetraacetic acid    -   EGTA Ethyleneglycol bis(2-aminoethyl ether)-N,N,N′,N′        tetraacetic acid    -   EtOAc Ethyl Acetate    -   EtOH Ethanol    -   HPLC-MS High Performance Liquid Chromatography Mass Spectrometry    -   HTS High Throughput Screen    -   HTRF Homogeneous Time-Resolved Fluorescence    -   LCMS Liquid Chromatography Mass Spectrometry    -   LiHMDS Lithium bis(trimethylsilyl)amide    -   MeOH Methanol    -   NIS N-Iodosuccinimide    -   RPMI-1640 A cell growth medium developed at Roswell Park        Memorial Institute    -   TBS t-Butyldimethylsilyl    -   TEA Triethylamine    -   TFA Trifluoroacetic acid    -   THF Tetrahydrofuran

Preparation of Examples Example 19-((1r,4r)-4-Methylcyclohexyl)-N-(5,6,7,8-tetrahydro-1,6-naphthyridin-2-yl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-amine

Synthesis of N⁴-((1r,4r)-4-methylcyclohexyl)pyridine-2,4-diamine

4-Chloropyrimidine-2-amine (commercially available from Sigma-Aldrich,St. Louis, Mo.) (1000 g, 7.72 mol, 1.0 eq),trans-4-methylcyclohexylamine hydrochloride (commercially available fromTCI America, M1780) (1500 g, 10.03 mol, 1.3 eq) and TEA (3.23 L, 23.2mol, 3.0 eq) were mixed together in n-butanol (8 L). The reactionmixture was heated at reflux for 36 hours and monitored using LCMS. Uponcompletion, the reaction mixture was cooled to room temperature, dilutedwith water (8 L) and extracted with EtOAc (2×10 L). The organic layerswere combined, dried over Na₂SO₄, and concentrated under reducedpressure to give the title compound (1770 g) which was used in the nextstep without further purification.

Synthesis of 5-iodo-N⁴-((1r,4r)-4-methylcyclohexyl)pyridine-2,4-diamine

N⁴-((1r,4r)-4-Methylcyclohexyl)pyridine-2,4-diamine (1770 g, 8.58 mol,1.0 eq) was dissolved in anhydrous DMF (8 L). To this solution under N₂atmosphere at 10° C. was added NIS (1.93 kg, 8.58 mol, 1.0 eq) inportions over 10 minutes. Upon completion of the addition, the reactionmixture was stirred at room temperature for 2 hours. The reaction wasmonitored using LCMS. Upon completion, the reaction mixture was cooledusing an ice bath, quenched with saturated aqueous sodium carbonate (5L) and extracted with EtOAc (2×15 L). The combined organic extracts werewashed with saturated aqueous sodium carbonate (2×5 L), water (3×2 L),dried over Na₂SO₄, and concentrated under reduced pressure. The residuewas purified using column chromatography eluting with 25% to 40% EtOAcin hexanes to provide the title compound (1.47 kg, 57% over two steps).¹H-NMR (300 MHz, DMSO-d₆) δ ppm 0.85 (3H, d, J=7.2 Hz), 0.98 (1H, dd,J=12.9, 2.7 Hz), 1.41-1.27 (3H, m), 1.66 (2H, d, J=12.3 Hz), 1.78 (2H,d, J=12.3 Hz), 3.85 (1H, m), 5.48 (1H, d, J=8.1 Hz), 6.16 (2H, br s),7.86 (1H, s) ppm; MS m/z: 333 (M+1).

Synthesis of5-(3-fluoropyridin-4-yl)-N⁴-((1r,4r)-4-methylcyclohexyl)pyrimidine-2,4-diamine

To a solution of 2,2,6,6-tetramethylpiperidine (commercially availablefrom Sigma-Aldrich, St. Louis, Mo.) (997 mL, 5.87 mol, 3 eq) inanhydrous THF (6 L) under N₂ atmosphere at 0° C., was added n-BuLi (2.5M in hexanes, 2.35 L, 5.87 mol, 3 eq) via an addition funnel over 30minutes. Upon completion of the addition, the reaction mixture wasstirred at 0° C. for 1 hour. The reaction mixture was cooled to −74° C.(acetone/dry ice bath) and a solution of 3-fluoropyridine (commerciallyavailable from Sigma-Aldrich, St. Louis, Mo.) (561 g, 5.773 mol, 2.95eq) in anhydrous THF (500 mL) was added over 15 minutes keeping thetemperature below −63° C. Upon completion of the addition, the reactionmixture was stirred at −74° C. for an additional 2 hours. A solution ofZnBr₂ (1422 g, 6.32 mol, 3.22 eq) in anhydrous THF (3 L) was then addeddropwise over 35 minutes keeping the temperature below −60° C. Uponcompletion of the addition, the cold bath was removed and the reactionmixture was allowed to warm to room temperature. Then5-iodo-N-((1r,4r)-4-methylcyclohexyl)pyridine-2,4-diamine (650 g, 1.95mol, 1.0 eq) was added in one portion followed by Pd(PPh₃)₄ (113 g, 97.8mmol, 0.05 eq). The reaction mixture was heated at reflux overnight andmonitored using LCMS. Upon completion, the reaction mixture was cooledto room temperature, quenched with saturated aqueous NaHCO₃ (6 L) andextracted with EtOAc (10 L×2). The organic extracts were washed withsaturated NaHCO₃ (2.5 L×2) and brine (2.5 L), and were then concentratedunder vacuum. The residue was dissolved in 2N HCl (2.5 L) and washedwith DCM (1.25 L×3). The aqueous phase was adjusted to pH 10-12 byaddition of aqueous 4N NaOH and extracted with DCM (1.5 L×3). Theorganic extracts were washed with water (1.25 L×2), dried andconcentrated to give the title compound (540 g, 92%). ¹H-NMR (300 MHz,DMSO-d₆) δ ppm 0.85 (3H, d, J=7.2 Hz), 0.98 (1H, dd, J=12.9, 2.7 Hz),1.30-1.18 (3H, m), 1.64 (2H, d, J=12.3 Hz), 1.74 (2H, d, J=11.7 Hz),3.96 (1H, m), 5.00 (1H, d, J=8.4 Hz), 6.24 (2H, br s), 7.35 (1H, dd,J=6.6, 4.4 Hz), 7.58 (1H, s), 8.37 (1H, d, J=4.8 Hz), 8.50 (1H, d, J=6.6Hz) ppm.

Synthesis of9-((1r,4r)-4-methylcyclohexyl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-amine.

To a solution of5-(3-fluoropyridin-4-yl)-N⁴-((1r,4r)-4-methylcyclohexyl)pyrimidine-2,4-diamine(854 g, 2.84 mol, 1.0 eq) in anhydrous 1-methyl-2-pyrrolidinone (8 L)under N₂ atmosphere at room temperature, was added LiHMDS (1.0 M intoluene, 8.5 L, 8.5 mol, 3.0 eq) over 30 minutes. Upon completion of theaddition, the reaction mixture was heated at 90° C. overnight andmonitored using LCMS. Upon completion, the reaction mixture was cooledto room temperature, quenched with ice cold water (10 L) and extractedwith EtOAc (12 L). The organic phase was washed with saturated aqueousNaHCO₃ (4 L×2), and water (2 L×3). The aqueous layers were combined andback extracted with EtOAc (15 L×2). The organic layers were combined,dried over Na₂SO₄, and concentrated under reduced pressure. The solidthus obtained was suspended in DCM (2.5 L) and agitated using a rotaryevaporator for 30 minutes. The solid was collected by filtration, washedwith DCM and dried to afford the title compound (400 g). The motherliquor was purified by column chromatography (eluting withDCM/MeOH=50:1) to afford, after triturating with DCM (750 mL),additional title compound (277 g, total: 677 g, yield: 84%). ¹H NMR (300MHz, CD₃OD) δ ppm 1.02 (d, J=6.3 Hz, 3H), 1.33-1.20 (m, 2H), 1.67-1.60(m, 2H), 1.95-1.84 (m, 4H), 1.58-1.45 (m, 2H), 4.87-4.77 (m, 1H), 7.94(d, J=5.1 Hz, 1H), 8.31 (d, J=5.1 Hz, 1H), 8.87 (s, 1H), 8.96 (s, 1H)ppm; MS m/z: 282.0 (M+1).

Synthesis of tert-butyl2-chloro-7,8-dihydro-1,6-naphthyridine-6(5H)-carboxylate

To a slurry of 2-chloro-5,6,7,8-tetrahydro-1,6-naphthyridinehydrochloride (106.1 g, 517 mmol, commercially available from D-L ChiralChemicals, ST-0143) and N,N-diisopropylethylamine (80 g, 108 mL, 621mmol, 1.2 eq) in DCM (1 L) was added a solution of di-tert-butyldicarbonate (119 g, 543 mmol, 1.05 eq) in DCM (100 mL) via an additionfunnel within 1 hr. The reaction mixture became a clean solution and thesolution thus obtained was stirred at room temperature for an additionalhour and monitored using LCMS. Upon completion, the reaction mixture wasconcentrated. The residue was dissolved in EtOAc (1 L) and washed withwater (3×300 mL), washed with brine (300 mL) and dried over Mg50₄. Thesolvent was evaporated under vacuum to give the title compound as anoff-white solid (139 g, yield: 100%). ¹H NMR (400 MHz, CDCl₃) δ ppm 1.49(9H, s), 2.97 (2H, t, J=5.9 Hz), 3.73 (2H, t, J=6.0 Hz), 4.57 (2H, s),7.17 (1H, d, J=8.0 Hz), 7.38 (1H, d, J=8.0 Hz) ppm; LCMS m/z: 269 (M+1).

Synthesis of tert-butyl2-((9-((1r,4r)-4-methylcyclohexyl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-yl)amino)-7,8-dihydro-1,6-naphthyridine-6(5H)-carboxylate

To a solution of9-((1r,4r)-4-methylcyclohexyl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-amine(2.81 g, 10 mmol) in 1,4-dioxane (45 mL) were added tert-butyl2-chloro-7,8-dihydro-1,6-naphthyridine-6(5H)-carboxylate (2.57 g, 9.55mmol), 4,5-bis(diphenylphosphino)-9,9-dimethylxanene (231 mg, 0.40mmol), and sodium t-butoxide (1.44 g, 15 mmol). Argon was bubbledthrough the mixture for 10 minutes.Tris(dibenzylideneacetone)dipalladium (0)(183 mg, 0.20 mmol) was added,and argon was again bubbled through the mixture for 5 minutes. Thereaction mixture thus obtained was stirred at 100° C. for 3 hourswhereupon HPLC-MS analysis indicated that the reaction was complete. Thereaction mixture was cooled to 40° C. and diluted with DCM (90 mL) andtreated with Si-triamine (functionalized silica gel, from Silicycle,FR31017TR130B) (2.8 g) overnight at room temperature. Celite® brandfilter aid 545 (6 g) was added, and the mixture was filtered with asintered glass funnel and the solid phase was rinsed with DCM (100 mL).The filtrate was concentrated to 25 mL on a rotary evaporator anddiluted with a mixture of EtOAc and hexane (20 mL, 4:1). The resultingslurry was stirred at room temperature for 5 hours. The solid wascollected by filtration, washed with a mixture of EtOAc and hexane (20mL, 1:1) and air dried for a few hours to provide the title compound asan off-white solid (4.90 g, 100% yield). ¹H NMR (500 MHz, CD₂Cl₂) δ ppm1.06 (3H, d, J=6.4 Hz), 1.34-1.22 (2H, m), 1.48 (9H, s), 1.67 (1H, br.s), 2.02-1.93 (4H, m), 2.63 (2H, dq, J=3.1, 12.8 Hz), 2.88 (2H, t, J=5.7Hz), 3.74 (2H, t, J=6.0 Hz), 4.57 (2H, s), 7.51 (1H, d, J=8.6 Hz), 7.85(1H, d, J=5.1 Hz), 8.10 (1H, br. s), 8.42 (1H, d, J=8.3 Hz), 8.46 (1H,d, J=4.9 Hz), 8.97 (1H, s), 9.10 (1H, s) ppm; LCMS m/z: 514(M+1).

Synthesis of9-((1r,4r)-4-methylcyclohexyl)-N-(5,6,7,8-tetrahydro-1,6-naphthyridin-2-yl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-amine(1).

To a suspension of tert-butyl2-((9-((1r,4r)-4-methylcyclohexyl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-yl)amino)-7,8-dihydro-1,6-naphthyridine-6(5H)-carboxylate:9-((1r,4r)-4-methylcyclohexyl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-amine(4.65 g, 9.05 mmol) in MeOH (30 mL) were added concentrated HCl (6.74mL) and water (14 mL). The mixture thus obtained was stirred at roomtemperature overnight. 50% NaOH in water (4.8 mL) was added at 0° C. tothe reaction mixture to adjust the pH value to 9. The precipitatedyellow solid was collected by filtration, rinsed with water (25 mL) andair dried for 3 days to give the title compound (3.75 g, 100%). ¹H NMR(400 MHz, CDCl₃) δ ppm 1.07 (3H, d, J=6.5 Hz), 1.29-1.25 (3H, m),2.00-1.95 (3H, m), 2.02 (2H, s), 2.69-2.53 (2H, m), 2.89 (2H, t, J=6.0Hz), 3.26 (2H, t, J=6.0 Hz), 4.04 (2H, s), 4.71 (1H, m, J=12.8, 12.8Hz), 7.41 (1H, d, J=8.4 Hz), 7.84 (1H, d, J=6.1 Hz), 7.84 (1H, d, J=6.1Hz), 8.03 (1H, s), 8.34 (1H, d, J=8.4 Hz), 8.50 (1H, d, J=5.3 Hz), 8.96(1H, s), 9.08 (1H, s) ppm; LCMS m/z: 414 (M+1).

Example 21-(2-((9-((1r,4r)-4-Methylcyclohexyl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-yl)amino)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)ethanone

Synthesis of1-(2-((9-((1r,4r)-4-methylcyclohexyl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-yl)amino)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)ethanone(2)

To a stirred ice-cooled solution of9-((1r,4r)-4-methylcyclohexyl)-N-(5,6,7,8-tetrahydro-1,6-naphthyridin-2-yl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-amine(1) (90 mg, 0.22 mmol) in 3 mL of 1,4-dioxane was added acetic anhydride(33.3 mg, 0.33 mmol). After stirring for 10 minutes, 20 mL of DCM and 10mL of water were added. The resulting solution was washed sequentiallywith saturated aqueous NaHCO₃ solution, water and brine, dried overanhydrous MgSO₄, and evaporated under vacuum. The residue was purifiedby preparatory LC to give the title compound (50 mg, 50%). ¹H NMR (400MHz, CD3OD) δ ppm 1.10 (3H, d, J=6.46 Hz) 1.31-1.43 (2H, m) 1.70-1.80(1H, br. s.) 2.00-2.09 (4H, m) 2.27 (3H, d, J=7.24 Hz) 2.67-2.79 (2H, m)3.10 (1H, t, J=6.16 Hz) 3.15-3.22 (1H, m) 3.98 (2H, dt, J=13.16, 6.04Hz) 4.79-4.84 (2H, m) 4.91-5.00 (1H, m) 7.90-7.98 (2H, m) 8.61-8.65 (2H,m) 9.40-9.41 (1H, m) 9.62 (1H, d, J=2.74 Hz) ppm; LCMS m/z: 456 (M+1).

Example 32-(2-((9-((1r,4r)-4-Methylcyclohexyl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-yl)amino)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)ethanol

Synthesis of2-(2-((9-((1r,4r)-4-methylcyclohexyl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-yl)amino)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)ethanol(3)

A slurry of9-((1r,4r)-4-methylcyclohexyl)-N-(5,6,7,8-tetrahydro-1,6-naphthyridin-2-yl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-amine(1) (50 mg, 0.12 mmol) and (tert-butyldimethylsilyloxy)acetaldehyde(commercially available from Sigma-Aldrich, St. Louis, Mo.) (31.6 mg,0.18 mmol) in 3 mL 1,4-dioxane was stirred for 5 minutes and then sodiumtriacetoxyborohydride (commercially available from Sigma-Aldrich, St.Louis, Mo.) (77 mg, 0.363 mmol) was added. After stirring for 10minutes, DCM (30 mL) was added to the reaction mixture. The resultingsolution was washed sequentially with saturated aqueous NaHCO₃ solution,water and brine, dried over anhydrous MgSO₄, and evaporated under vacuumto give the intermediate TBS protected alcohol (LCMS m/z: 572 (M+1)).The TBS protected alcohol was treated with 3 mL of 4 N HCl/dioxane toremove the TBS group. After 30 minutes, the reaction was concentratedand purified by flash chromatography on silica gel eluting with 0 to 20%2N NH₃ MeOH solution in DCM to give the title compound (38 mg, 68%yield). ¹H NMR (400 MHz, CDCl₃) δ ppm 1.09 (3H, d, J=6.65 Hz) 1.24-1.36(2H, m) 1.56-1.73 (3H, m) 1.96-2.06 (4H, m) 2.63 (2H, qd, J=12.88, 3.81Hz) 2.81 (2H, t, J=5.38 Hz) 2.94-3.04 (4H, m) 3.73-3.80 (4H, m)4.70-4.79 (1H, m) 7.44 (1H, m, J=8.61 Hz) 7.86 (1H, dd, J=5.18, 1.08 Hz)8.08 (1H, s) 8.37 (1H, m, J=8.41 Hz) 8.53 (1H, d, J=5.28 Hz) 8.97-9.00(1H, m) 9.10-9.12 (1H, m) ppm; LC/MS m/z: 458 (M+1).

Example 43-(2-((9-((1r,4r)-4-Methylcyclohexyl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-yl)amino)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)propan-1-ol

Synthesis of3-(2-((9-((1r,4r)-4-methylcyclohexyl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-yl)amino)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)propan-1-ol(4)

The title compound (4) was prepared from9-((1r,4r)-4-methylcyclohexyl)-N-(5,6,7,8-tetrahydro-1,6-naphthyridin-2-yl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-amine(1) using chemistry similar to that described in Example 3 using(tert-butyldimethylsilyloxy)propanal (commercially available fromChemPacific, Baltimore, Md.) in place of(tert-butyldimethylsilyloxy)acetaldehyde. ¹H NMR (400 MHz, CD₃OD) δ ppm1.10 (3H, d, J=6.65 Hz), 1.32-1.44 (2H, m), 1.75 (1H, m), 1.98-2.16 (6H,m), 2.37-2.45 (2H, m), 2.67-2.80 (2H, m), 3.30-3.40 (2H, m), 3.46-3.57(2H, m), 3.66-3.90 (2H, m), 4.55-4.64 (2H, m), 4.95 (1H, m), 7.81 (1H,dd, J=8.51, 1.27 Hz), 8.36 (1H, d, J=8.80 Hz), 8.62 (2H, q, J=6.26 Hz),9.42 (1H, s), 9.59 (1H, m) ppm; LC/MS m/z: 472 (M+1).

Example 52-Hydroxy-1-(2-β9-((1r,4r)-4-methylcyclohexyl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-yl)amino)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)ethanone

Synthesis of 2,5-dioxopyrrolidin-1-yl 2-acetoxyacetate

A 3-neck round-bottom flask equipped with a mechanical stirrer,thermocouple and addition funnel with nitrogen inlet was charged withN-hydroxysuccinimide (commercially available from Sigma-Aldrich, St.Louis, Mo.) (211 g, 1.83 mol) and DCM (2.25 L) at room temperature,resulting in a suspension. Pyridine (178 mL, 2.2 mol) was added in oneportion with no change in the internal temperature. A solution ofacetoxyacetyl chloride (commercially available from Sigma-Aldrich, St.Louis, Mo.) (197 mL, 1.83 mol) in DCM (225 mL) was added dropwise over60 minutes and the temperature rose to 35° C. Stirring was continued atroom temperature for 2.5 hours. The reaction mixture was washed withwater (1×1 L), 1N HCl (2×1 L) and brine (1×1 L). The organic layer wasconcentrated under vacuum and azeotroped with toluene (1×1 L) to obtainthe product as a white solid (367 g, 93%). ¹H NMR (400 MHz, CDCl₃) δ4.96 (2H, s), 2.86 (4H, s), 2.19 (3H, s) ppm; LCMS m/z: 238 (M+Na).

Synthesis of2-(2-((9-((1r,4r)-4-methylcyclohexyl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-yl)amino)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)-2-oxoethylacetate

To a suspension of9-((1r,4r)-4-methylcyclohexyl)-N-(5,6,7,8-tetrahydro-1,6-naphthyridin-2-yl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-amine(1) (827 mg, 2.0 mmol) in chloroform (10 mL) were addeddiisopropylethylamine (258 mg, 348 uL, 2.0 mmol) and2,5-dioxopyrrolidin-1-yl2-acetoxyacetate (560 mg, 2.6 mmol). Thereaction mixture thus obtained was stirred at room temperature for 30minutes whereupon the mixture became a yellow solution. HPLC-MS analysisindicated that the reaction was complete. The reaction mixture wasconcentrated. MeOH (5 mL) and water (6 mL) were added to form a slurrywhich was stirred at room temperature for 1 hour. The solid wascollected by filtration to give the title compound as a light yellowsolid (1.04 g, 98% yield). ¹H NMR (400 MHz, CDCl₃, rotamers) δ ppm 1.08(3H, d, J=6.5 Hz), 1.37-1.20 (2H, m), 2.03-1.97 (4H, m), 2.22 (3H, s),2.69-2.52 (2H, m, J=2.9, 12.8, 12.8, 12.8 Hz), 3.08-2.93 (2H, m), 3.75(1H, t, J=5.9 Hz), 3.97 (1H, t, J=5.6 Hz), 4.59 (1H, s),), 4.80-4.65(2H, m),), 4.90-4.82 (2H, m), 7.57-7.45 (1H, m), 7.86 (1H, d, J=5.7 Hz),8.21-8.10 (1H, m), 8.49-8.40 (1H, m), 8.52 (1H, d, J=5.3 Hz), 8.98 (1H,s), 9.11 (1H, s) ppm; LCMS m/z: 514 (M+1).

Synthesis of2-hydroxy-1-(2-((9-((1r,4r)-4-methylcyclohexyl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-yl)amino)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)ethanone(5)

To a solution of2-(2-((9-((1r,4r)-4-methylcyclohexyl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-yl)amino)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)-2-oxoethylacetate (514 mg, 1.0 mmol) in DCM (7.5 mL) and MeOH (2.5 mL) was added0.5 M sodium methoxide solution in MeOH (0.30 mL, 0.15 mmol), and thereaction mixture was stirred at room temperature for 1 hour andmonitored using LCMS. Upon completion, the reaction mixture wasconcentrated. The residue was treated with EtOH (5 mL) and water (10 mL)to provide a solid which was collected by filtration, washed with water,and dried in a vacuum oven at 55° C. overnight to give the titlecompound (5) as a white solid (468 mg, 99% yield). ¹H NMR (500 MHz,acetic acid-d₄, 373 K) δ ppm 1.09 (3H, d, J=6.5 Hz), 1.31-1.43 (2H, m),1.70-1.80 (1H, m), 1.99-2.03 (2H, m), 2.06-2.13 (2H, m), 2.68 (2H, dq,J=3.3, 12.7 Hz), 3.10 (2H, t, J=5.4 Hz), 3.88 (2H, br. s.), 4.46 (2H,br. s.), 4.77 (2H, br. s), 4.90 (1H, tt, J=3.9, 12.4 Hz), 7.76 (1H, d,J=8.5 Hz), 8.33 (1H, d, J=8.5 Hz), 8.40 (1H, d, J=6.0 Hz), 8.63 (1H, d,J=6.0 Hz), 9.35 (1H, s), 9.43 (1H, s) ppm; LCMS m/z: 472 (M+1).

Synthesis of2-hydroxy-1-(2-((9-((1r,4r)-4-methylcyclohexyl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-yl)amino)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)ethanonemonohydrochloride dihydrate

To a suspension of2-hydroxy-1-(2-((9-((1r,4r)-4-methylcyclohexyl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-yl)amino)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)ethanone(472 mg, 1.0 mmol) in water (2 mL) was added 2 N HCl (2 mL). The mixturebecame a clear solution. The pH value of the solution was adjusted to 4by addition of 2 N NaOH at 0° C. and the precipitated light yellow solidwas collected by filtration. The collected solid was washed with coldwater three times. The solid was dried under vacuum to give the titlecompound as a light yellow solid (469 mg, 92% yield). ¹H NMR (500 MHz,DMSO-d₆) δ 1.02 (3H, d, J=5.0 Hz), 1.20-1.30 (2H, m), 1.64 (1H, m),1.88-1.90 (4H, m), 2.59-2.66 (2H, m), 2.85-2.95 (2H, m), 3.71 (1H, m),3.83 (1H, m), 4.19-4.22 (2H, m), 4.60-4.67 (2H, m), 4.85 (1H, m), 7.75(1H, d, J=8.5 Hz), 8.19 (1H, d, J=8.5 Hz), 8.55 (1H, d, J=5.0 Hz), 8.63(1H, d, J=5.0 Hz), 9.47 (1H, s), 9.58 (1H, s), 10.59 (1H, br.s) ppm;LCMS m/z: 472 (M+1). Anal. (C₂₆H₂₉N₇O₂.HCl.2H₂O) Calc: C=57.40, H=6.30,N=18.02. Found: C=57.06, H=6.31, N=17.92.

Alternative Synthesis of Hydrochloride Salt of2-Hydroxy-1-(2-((9-((1r,4r)-4-methylcyclohexyl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-yl)amino)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)ethanone

To a suspension of2-hydroxy-1-(2-((9-((1r,4r)-4-methylcyclohexyl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-yl)amino)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)ethanone(2.385 g, 5.0 mmol) in water (10 mL) was added 2N HCl (10 mL) at 20° C.The mixture became a clear light yellow solution. The pH value of thesolution was adjusted to 4 by addition of 2N NaOH through additionfunnel at 0° C., and the precipitated yellow solid was collected byfiltration. The resulting solid was washed with cold water three times.The solid was dried under vacuum at 50° C. for two days to provide 2.49g of the hydrochloride salt of2-hydroxy-1-(2-((9-((1r,4r)-4-methylcyclohexyl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-yl)amino)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)ethanoneas a solid. This salt was also obtained as a hydrate.

Preparation of Other Salts of Example 5

The following salts were prepared as described below. Whether or notthese were obtained as the hydrates or as anhydrous forms has not yetbeen determined.

Synthesis of Phosphate Salt of2-Hydroxy-1-(2-((9-((1r,4r)-4-methylcyclohexyl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-yl)amino)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)ethanone

2-Hydroxy-1-(2-((9-((1r,4r)-4-methylcyclohexyl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-yl)amino)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)ethanone(100 mg) was added to a 20 mL vial. Approximately 15 mL of a mixture of70% DCM and 30% EtOH (v/v) was added to the vial, and the mixture wasvortexed for approximately 30 seconds to dissolve most of the neutralcompound. Aqueous 0.1 M phosphoric acid (1 equivalent) was added to thevial, and the mixture was sonicated for approximately 30 seconds. Thesolvents were then allowed to evaporate slowly to provide the phosphatesalt of2-hydroxy-1-(249-((1r,4r)-4-methylcyclohexyl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-yl)amino)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)ethanone.The salt was characterized using XRPD and differential scanningcalorimetry thermograms.

Synthesis of Citrate Salt of2-Hydroxy-1-(2-((9-((1r,4r)-4-methylcyclohexyl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-yl)amino)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)ethanone

2-Hydroxy-1-(2-((9-((1r,4r)-4-methylcyclohexyl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-yl)amino)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)ethanone(100 mg) was added to a 20 mL vial. Approximately 15 mL of a mixture of70% DCM and 30% EtOH (v/v) was added to the vial, and the mixture wasvortexed for approximately 30 seconds to dissolve most of the neutralcompound. Aqueous 0.1 M citric acid (1 equivalent) was added to thevial, and the mixture was sonicated for approximately 30 seconds. Thesolvents were then allowed to evaporate slowly to provide the citratesalt of2-hydroxy-1-(2-((1r,4r)-4-methylcyclohexyl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-yl)amino)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)ethanone.The salt was characterized using XRPD and differential scanningcalorimetry thermograms.

Synthesis of Tartrate Salt of2-Hydroxy-1-(2-((9-((1r,4r)-4-methylcyclohexyl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-yl)amino)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)ethanone

2-Hydroxy-1-(2-((9-((1r,4r)-4-methylcyclohexyl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-yl)amino)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)ethanone(100 mg) was added to a 20 mL vial. Approximately 15 mL of a mixture of70% DCM and 30% EtOH (v/v) was added to the vial, and the mixture wasvortexed for approximately 30 seconds to dissolve most of the neutralcompound. Aqueous 0.1 M tartaric acid (1 equivalent) was added to thevial, and the mixture was sonicated for approximately 30 seconds. Thesolvents were then allowed to evaporate slowly to provide the tartratesalt of2-hydroxy-1-(2-((9-((1r,4r)-4-methylcyclohexyl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-yl)amino)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)ethanone.The salt was characterized using XRPD and differential scanningcalorimetry thermograms.

Synthesis of Salicylate Salt of2-Hydroxy-1-(2-((9-((1r,4r)-4-methylcyclohexyl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-yl)amino)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)ethanone

2-Hydroxy-1-(2-((9-((1r,4r)-4-methylcyclohexyl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-yl)amino)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)ethanone(100 mg) was added to a 20 mL vial. Approximately 15 mL of a mixture of70% DCM and 30% EtOH (v/v) was added to the vial, and the mixture wasvortexed for approximately 30 seconds to dissolve most of the neutralcompound. Aqueous 0.1 M salicylic acid (1 equivalent) was added to thevial, and the mixture was sonicated for approximately 30 seconds. Thesolvents were then allowed to evaporate slowly to provide the salicylatesalt of2-hydroxy-1-(2-((9-((1r,4r)-4-methylcyclohexyl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-yl)amino)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)ethanone.The salt was characterized using XRPD and differential scanningcalorimetry thermograms.

Synthesis of Benzenesulfonate (Besylate) Salt of2-Hydroxy-1-(2-((9-((1r,4r)-4-methylcyclohexyl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-yl)amino)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)ethanone

2-Hydroxy-1-(2-((9-((1r,4r)-4-methylcyclohexyl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-yl)amino)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)ethanone(100 mg) was added to a 20 mL vial. Approximately 15 mL of a mixture of70% DCM and 30% EtOH (v/v) was added to the vial, and the mixture wasvortexed for approximately 30 seconds to dissolve most of the neutralcompound. Aqueous 0.1 M benzenesulfonic acid (1 equivalent) was added tothe vial, and the mixture was sonicated for approximately 30 seconds.The solvents were then allowed to evaporate slowly to provide thebenzenesulfonate salt of2-hydroxy-1-(2-((9-((1r,4r)-4-methylcyclohexyl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-yl)amino)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)ethanone.The salt was characterized using XRPD and differential scanningcalorimetry thermograms.

Synthesis of p-Toluenesulfonate (Tosylate) Salt of2-Hydroxy-1-(2-((9-((1r,4r)-4-methylcyclohexyl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-yl)amino)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)ethanone

2-Hydroxy-1-(2-((9-((1r,4r)-4-methylcyclohexyl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-yl)amino)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)ethanone(100 mg) was added to a 20 mL vial. Approximately 15 mL of a mixture of70% DCM and 30% EtOH (v/v) was added to the vial, and the mixture wasvortexed for approximately 30 seconds to dissolve most of the neutralcompound. Aqueous 0.1 M p-toluenesulfonic acid (1 equivalent) was addedto the vial, and the mixture was sonicated for approximately 30 seconds.The solvents were then allowed to evaporate slowly to provide thetosylate salt of2-hydroxy-1-(2-((9-((1r,4r)-4-methylcyclohexyl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-yl)amino)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)ethanone.The salt was characterized using XRPD and differential scanningcalorimetry thermograms.

Synthesis of ethanesulfonate Salt of2-Hydroxy-1-(2-((9-((1r,4r)-4-methylcyclohexyl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-yl)amino)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)ethanone

2-Hydroxy-1-(2-((9-((1r,4r)-4-methylcyclohexyl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-yl)amino)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)ethanone(100 mg) was added to a 20 mL vial. Approximately 15 mL of a mixture of70% DCM and 30% EtOH (v/v) was added to the vial, and the mixture wasvortexed for approximately 30 seconds to dissolve most of the neutralcompound. Aqueous 0.1 M ethanesulfonic acid (1 equivalent) was added tothe vial, and the mixture was sonicated for approximately 30 seconds.The solvents were then allowed to evaporate slowly to provide theethanesulfonate salt of2-hydroxy-1-(2-((9-((1r,4r)-4-methylcyclohexyl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-yl)amino)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)ethanone.The salt was characterized using XRPD and differential scanningcalorimetry thermograms.

Synthesis of Sulfate Salt (1 Equivalent) of2-Hydroxy-1-(2-((9-((1r,4r)-4-methylcyclohexyl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-yl)amino)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)ethanone

2-Hydroxy-1-(2-((9-((1r,4r)-4-methylcyclohexyl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-yl)amino)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)ethanone(100 mg) was added to a 20 mL vial. Approximately 15 mL of a mixture of70% DCM and 30% EtOH (v/v) was added to the vial, and the mixture wasvortexed for approximately 30 seconds to dissolve most of the neutralcompound. Aqueous 0.1 M sulfuric acid (1 equivalent) was added to thevial, and the mixture was sonicated for approximately 30 seconds. Thesolvents were then allowed to evaporate slowly to provide the sulfatesalt (1 equivalent) of2-hydroxy-1-(2-((9-((1r,4r)-4-methylcyclohexyl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-yl)amino)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)ethanone.The salt was characterized using XRPD and differential scanningcalorimetry thermograms.

Synthesis of Sulfate Salt (½ Equivalent) of2-Hydroxy-1-(2-((9-((1r,4r)-4-methylcyclohexyl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-yl)amino)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)ethanone

2-Hydroxy-1-(2-((9-((1r,4r)-4-methylcyclohexyl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-yl)amino)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)ethanone(100 mg) was added to a 20 mL vial. Approximately 15 mL of a mixture of70% DCM and 30% EtOH (v/v) was added to the vial, and the mixture wasvortexed for approximately 30 seconds to dissolve most of the neutralcompound. Aqueous 0.1 M sulfuric acid (½ equivalent) was added to thevial, and the mixture was sonicated for approximately 30 seconds. Thesolvents were then allowed to evaporate slowly to provide the sulfatesalt (½ equivalent) of2-hydroxy-1-(2-((9-((1r,4r)-4-methylcyclohexyl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-yl)amino)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)ethanone.The salt was characterized using XRPD and differential scanningcalorimetry thermograms.

Synthesis of Ethanedisulfonate (1 Equivalent) Salt of2-Hydroxy-1-(2-((9-((1r,4r)-4-methylcyclohexyl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-yl)amino)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)ethanone

2-Hydroxy-1-(2-((9-((1r,4r)-4-methylcyclohexyl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-yl)amino)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)ethanone(100 mg) was added to a 20 mL vial. Approximately 15 mL of a mixture of70% DCM and 30% EtOH (v/v) was added to the vial, and the mixture wasvortexed for approximately 30 seconds to dissolve most of the neutralcompound. Aqueous 0.1 M ethanedisulfonic acid (1 equivalent) was addedto the vial, and the mixture was sonicated for approximately 30 seconds.The solvents were then allowed to evaporate slowly to provide theethanedisulfonate (1 equivalent) salt of2-hydroxy-1-(2-((9-((1r,4r)-4-methylcyclohexyl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-yl)amino)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)ethanone.The salt was characterized using XRPD and differential scanningcalorimetry thermograms.

Synthesis of Ethanedisulfonate (½ Equivalent) Salt of2-Hydroxy-1-(2-[((9-((1r,4r)-4-methylcyclohexyl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-yl]amino)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)ethanone

2-Hydroxy-1-(2-((9-((1r,4r)-4-methylcyclohexyl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-yl)amino)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)ethanone(100 mg) was added to a 20 mL vial. Approximately 15 mL of a mixture of70% DCM and 30% EtOH (v/v) was added to the vial, and the mixture wasvortexed for approximately 30 seconds to dissolve most of the neutralcompound. Aqueous 0.1 M ethanedisulfonic acid (½ equivalent) was addedto the vial, and the mixture was sonicated for approximately 30 seconds.The solvents were then allowed to evaporate slowly to provide theethanedisulfonate (½ equivalent) salt of2-hydroxy-1-(2-((9-((1r,4r)-4-methylcyclohexyl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-yl)amino)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)ethanone.The salt was characterized using XRPD and differential scanningcalorimetry thermograms.

Example 62-(2-((9-((1r,4r)-4-Methylcyclohexyl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-yl)amino)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)-2-oxoaceticacid

Synthesis of methyl2-((6-(2-methoxy-2-oxoacetyl)-5,6,7,8-tetrahydro-1,6-naphthyridin-2-yl)(9-((1r,4r)-4-methylcyclohexyl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-yl)amino)-2-oxoacetate.

To a solution at room temperature of9-((1r,4r)-4-methylcyclohexyl)-N-(5,6,7,8-tetrahydro-1,6-naphthyridin-2-yl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-amine(1) (83 mg, 0.2 mmol) in THF (10 mL), were added pyridine (129 uL, 1.6mmol) and methyl chlorooxoacetate (commercially available fromSigma-Aldrich, St. Louis, Mo.) (74 uL, 0.8 mmol). The mixture thusobtained was stirred at room temperature overnight. The reaction mixturewas concentrated to give the title compound which was used in the nextstep without purification. LCMS m/z: 586 (M+1).

Synthesis of2-(2-((9-((1r,4r)-4-methylcyclohexyl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-yl)amino)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)-2-oxoaceticacid (6)

To a solution of unpurified methyl2-((6-(2-methoxy-2-oxoacetyl)-5,6,7,8-tetrahydro-1,6-naphthyridin-2-yl)(9-((1r,4r)-4-methylcyclohexyl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-yl)amino)-2-oxoacetatein THF/MeOH/H₂O (3/1/1, 5 mL) was added lithium hydroxide (19 mg, 0.80mmol). The mixture thus obtained was stirred at room temperature for 2hours. The reaction mixture was concentrated, and the residue wasdissolved in acidic water and purified by reverse phase prep-HPLC. Thefractions containing the desired product were collected and lyophilizedto give the title compound (6) as a light yellow solid (32 mg, 33% overtwo steps). ¹H NMR (500 MHz, DMSO-d₆) δ ppm 1.02 (3H, d, J=5.0 Hz), 1.28(2H, m), 1.65 (1H, m), 1.85-1.95 (4H, m), 2.60-2.70 (2H, m), 2.91 (1H,m), 2.95 (2H, m), 3.77 (2H, m), 4.70 (1H, s), 4.84 (1H, m), 7.79 (1H, d,J=10 Hz), 8.21 (1H, dd, J₁=10 Hz, J₂=5.0 Hz), 8.59 (1H, m), 8.66 (1H, d,J=5.0 Hz), 9.47 (1H, br. s), 9.59 (1H, s), 10.65 (1H, br. s) ppm; LCMSm/z: 486 (M+1).

Example 79-((1r,4r)-4-Methylcyclohexyl)-N-(6-(3,3,3-trifluoropropyl)-5,6,7,8-tetrahydro-1,6-naphthyridin-2-yl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-amine

Synthesis of9-((1r,4r)-4-methylcyclohexyl)-N-(6-(3,3,3-trifluoropropyl)-5,6,7,8-tetrahydro-1,6-naphthyridin-2-yl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-amine(7).

A slurry of9-((1r,4r)-4-methylcyclohexyl)-N-(5,6,7,8-tetrahydro-1,6-naphthyridin-2-yl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-amine(1) (40 mg, 0.10 mmol) and 3,3,3-trifluoropropanal (commerciallyavailable from ChemPacific, Baltimore, Md.) (10.8 mg, 0.10 mmol) in1,4-dioxane (2 mL) was stirred for 5 minutes, and then sodiumtriacetoxyborohydride (61.5 mg, 0.29 mmol) was added. After 10 minutes,the reaction mixture was diluted with DCM (30 mL). The resultingsolution was washed sequentially with saturated aqueous NaHCO₃ solution,water and brine, dried over anhydrous MgSO₄, and concentrated undervacuum. The residue was purified by preparative LC to give the titlecompound (7) (32 mg, 65%). ¹H NMR (400 MHz, CD₃OD) δ ppm 1.10 (3H, d,J=6.46 Hz), 1.31-1.45 (2H, m), 1.69-1.81 (1H, m), 1.98-2.09 (4H, m),2.67-2.79 (2H, m), 2.90-3.03 (2H, m), 3.62-3.70 (2H, m), 3.80 (2H, t,J=6.36 Hz), 4.58 (2H, s), 4.92-5.00 (3H, m), 7.80 (1H, d, J=8.61 Hz),8.36 (1H, d, J=8.80 Hz), 8.62 (2H, q, J=6.13 Hz), 9.41 (1H, s), 9.58(1H, s) ppm; LC/MS m/z: 510 (M+1).

Example 89-((1r,4r)-4-Methylcyclohexyl)-N-(6-(2-(methylsulfonyl)ethyl)-5,6,7,8-tetrahydro-1,6-naphthyridin-2-yl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-amine

Synthesis of2-chloro-6-(2-(methylsulfonyl)ethyl)-5,6,7,8-tetrahydro-1,6-naphthyridine

Methylsulfonylethene (commercially available from Sigma-Aldrich, St.Louis, Mo.) (142 mg, 1.33 mmol) was added dropwise to a solution of2-chloro-5,6,7,8-tetrahydro-1,6-naphthyridine (150 mg, 0.89 mmol,commercially available from D-L Chiral Chemicals, ST-0143) in EtOH (10mL). The reaction mixture was stirred at 60° C. for 2 hours, and thenconcentrated and purified by flash chromatography on silica gel elutingwith 0% to 5% MeOH in DCM to give2-chloro-6-(2-(methylsulfonyl)ethyl)-5,6,7,8-tetrahydro-1,6-naphthyridine(220 mg, 90% yield). ¹H NMR (400 MHz, CD₃OD) δ ppm 2.91-3.11 (9H, m)3.43 (2H, t, J=6.65 Hz) 3.73-3.77 (2H, m) 7.26 (1H, d, J=8.22 Hz) 7.56(1H, d, J=8.02 Hz) ppm; LC/MS m/z: 275 (M+1).

Synthesis of9-((1r,4r)-4-methylcyclohexyl)-N-(6-(2-(methylsulfonyl)ethyl)-5,6,7,8-tetrahydro-1,6-naphthyridin-2-yl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-amine(8).

The title compound was prepared using chemistry similar to thatdescribed in Example 1 using2-chloro-6-(2-(methylsulfonyl)ethyl)-5,6,7,8-tetrahydro-1,6-naphthyridinein place of tert-butyl2-chloro-7,8-dihydro-1,6-naphthyridine-6(5H)-carboxylate. ¹H NMR (400MHz, DMSO-d₆) δ ppm 1.09 (3H, d, J=6.60 Hz), 1.34 (1H, br. s.),1.26-1.40 (2H, m), 1.68-1.72 (1H, br. s.), 1.98-2.04 (4H, m), 2.66-2.72(2H, m), 2.98-3.06 (4H, m), 3.09 (3H, s), 3.13 (2H, t, J=6.60 Hz),3.43-3.48 (2H, m), 3.80 (2H, s), 4.89 (1H, br. s.), 7.62 (1H, d, J=8.31Hz), 8.14-8.15 (1H, m), 8.25 (1H, d, J=8.22 Hz), 8.45 (1H, d, J=5.38Hz), 9.05 (1H, s), 9.28-9.29 (1H, m) ppm; LC/MS m/z: 520 (M+1).

Example 9N-(6-(2-Fluoroethyl)-5,6,7,8-tetrahydro-1,6-naphthyridin-2-yl)-9-((1r,4r)-4-methylcyclohexyl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-amine

Synthesis ofN-(6-(2-fluoroethyl)-5,6,7,8-tetrahydro-1,6-naphthyridin-2-yl)-9-((1r,4r)-4-methylcyclohexyl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-amine(9)

To a solution of9-((1r,4r)-4-methylcyclohexyl)-N-(5,6,7,8-tetrahydro-1,6-naphthyridin-2-yl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-amine(1) (149 mg, 0.36 mmol) in absolute EtOH (4 mL) and DMF (2 mL), wasadded 1-bromo-2-fluoroethane (commercially available from AK Scientific,Mountain View, Calif.) (50 μL, 0.67 mmol). The resulting reactionmixture was stirred while being heated at 90° C. in a pre-heated oilbath. After 70 minutes of heating, a second aliquot of1-bromo-2-fluoroethane (50 μL, 0.67 mmol) was added. After 7.5 hours ofheating, when LCMS analysis indicated that the majority of the startingmaterial had been consumed, the mixture was allowed to cool andvolatiles were removed in vacuo. The resulting solution was diluted inDCM and sequentially washed with aqueous saturated Na₂CO₃, water, andbrine. The organic layers were dried over sodium sulfate, filtered, andconcentrated to a residue which was purified by flash chromatography onsilica gel eluting with a 10% to 45% gradient of solvent A(DCM:MeOH:NH₄OH, 90:9:1) in DCM. Chromatography fractions containing thedesired product were combined and stripped of solvents to provide aresidue which was further purified by precipitation from DCM/hexanes.The suspension so obtained was sonicated and filtered. The filter cakewas dried under high vacuum overnight to give the title compound (9) asan off-white solid (35 mg, 21% yield). ¹H NMR (400 MHz, CDCl₃): δ ppm1.07 (3H, d, J=6.5 Hz), 1.33-1.22 (2H, m), 1.85-1.69 (3H, br. m),2.00-1.96 (4H, m), 2.66-2.56 (2H, m), 3.02-2.90 (6H, m), 3.78 (2H, s),4.71 (2H, dt, J_(F,H)=47.6 Hz, J_(H,H)=4.8 Hz), 4.72 (1H, m), 7.42 (1H,d, J=8.4 Hz), 7.84 (1H, dd, J₁=5.1 Hz, J₂=0.98 Hz), 8.12 (1H, br. s),8.35 (1H, d, J=8.4 Hz), 8.50 (1H, d, J=5.3 Hz), 8.96 (1H, s), 9.10 (1H,s) ppm; MS m/z: 460.4 (M+1), 482.2 (M+Na).

Example 10N-(6-(2-Methoxyethyl)-5,6,7,8-tetrahydro-1,6-naphthyridin-2-yl)-9-((1r,4r)-4-methylcyclohexyl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-amine

Synthesis ofN-(6-(2-methoxyethyl)-5,6,7,8-tetrahydro-1,6-naphthyridin-2-yl)-9-((1r,4r)-4-methylcyclohexyl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-amine(10)

An aqueous solution of methoxyacetaldehyde of indeterminateconcentration was obtained by heating a solution of1,1,2-trimethoxyethane (commercially available from Sigma-Aldrich, St.Louis, Mo.) (10 g, 83 mmol) in 0.5 M HCl (150 mL) at 50° C. for 30minutes, followed by fractional distillation at atmospheric pressure.Sodium triacetoxyborohydride (331 mg, 1.56 mmol) was added to a solutionof9-((1r,4r)-4-methylcyclohexyl)-N-(5,6,7,8-tetrahydro-1,6-naphthyridin-2-yl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-amine(1) (101 mg, 0.24 mmol) in DCM (15 mL) containing glacial AcOH (100 μL,1.7 mmol) and 400 μL of the methoxyacetaldehyde solution obtained above.The reaction mixture was stirred at ambient temperature, and additionalsodium triacetoxyborohydride (474 mg, 2.24 mmol, in two successiveportions) and methoxyacetaldehyde (100 μL of the aqueous solution) wereadded until LCMS analysis indicated that the starting material had beencompletely consumed. The reaction was then quenched by addition of MeOH(6 mL), and stripped of volatiles on a rotary evaporator. The resultingresidue was purified by flash chromatography on silica gel eluting witha 10% to 60% gradient of solvent A (DCM:MeOH:NH₄OH; 90:9:1) in DCM.Chromatography fractions containing the desired product were combinedand concentrated in vacuo to give a residue that was further purified byprecipitation from DCM/hexanes. The suspension obtained in this way wassonicated and vacuum filtered. The filter cake was dried under highvacuum overnight to provide the titled compound (10) (32.6 mg, 28%yield). ¹H NMR (400 MHz, CDCl₃): δ ppm 1.07 (3H, d, J=6.5 Hz), 1.33-1.23(2H, m), 1.65 (3H, br. s), 2.01-1.96 (4H, m), 2.66-2.56 (2H, m),2.86-2.83 (2H, m), 3.02-2.97 (4H, m), 3.42 (3H, s), 3.67 (2H, t, J=5.3Hz), 3.76 (2H, br. s), 4.76-4.68 (1H, m), 7.41 (1H, d, J=8.6 Hz), 7.83(1H, dd, J₁=5.3 Hz, J₂=0.98 Hz), 8.08 (1H, br. s), 8.34 (1H, d, J=8.4Hz), 8.50 (1H, d, J=5.3 Hz), 8.96 (1H, s), 9.09 (1H, s) ppm; MS m/z:472.3 (M+1), 494.2 (M+Na).

Example 119-((1r,4r)-4-Methylcyclohexyl)-N-(5,6,7,8-tetrahydro-2,6-naphthyridin-3-yl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-amine

Synthesis of9-((1r,4r)-4-methylcyclohexyl)-N-(5,6,7,8-tetrahydro-2,6-naphthyridin-3-yl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-amine(11)

The title compound was prepared using chemistry similar to thatdescribed in Example 1 using7-chloro-1,2,3,4-tetrahydro-2,6-naphthyridine hydrochloride(commercially available from Anichem, North Brunswick, N.J.) in place of2-chloro-5,6,7,8-tetrahydro-1,6-naphthyridine hydrochloride. ¹H NMR (400MHz, CD₃OD) δ ppm 1.10 (3H, d, J=6.46 Hz), 1.33-1.45 (2H, m), 1.80-1.70(1H, m), 2.01-2.10 (4H, m), 2.68-2.77 (2H, m), 3.22 (2H, t, J=6.36 Hz),3.64 (2H, t, J=6.36 Hz), 4.93-5.02 (1H, m), 8.04 (1H, s), 8.39 (1H, s),8.63-8.70 (2H, m), 9.45 (1H, s), 9.63 (1H, s) ppm; LC/MS m/z: 414 (M+1).

Example 122-(7-((9-((1r,4r)-4-Methylcyclohexyl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-yl)amino)-3,4-dihydro-2,6-naphthyridin-2(1H)-yl)ethanol

Synthesis of2-(7-((9-((1r,4r)-4-methylcyclohexyl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-yl)amino)-3,4-dihydro-2,6-naphthyridin-2(1H)-yl)ethanol(12)

The title compound was prepared using chemistry similar to thatdescribed in Example 3 using9-((1r,40-4-methylcyclohexyl)-N-(5,6,7,8-tetrahydro-2,6-naphthyridin-3-yl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-amine(11) in place of9-((1r,4r)-4-methylcyclohexyl)-N-(5,6,7,8-tetrahydro-1,6-naphthyridin-2-yl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-amine(1). ¹H NMR (500 MHz, CD₃OD) δ ppm 1.11 (3H, d, J=6.60 Hz), 1.28-1.46(2H, m), 1.7-1.8 (1H, m) 1.86-2.09 (4H, m), 2.61-2.76 (2H, m), 2.79 (2H,t, J=5.99 Hz), 2.86-3.01 (4H, m), 3.8-3.9 (4H, m), 4.98 (1H, m),8.05-8.09 (1H, m), 8.11 (1H, s), 8.26-8.30 (1H, m), 8.39-8.44 (1H, m),8.98-9.03 (1H, m), 9.21-9.25 (1H, m) ppm; LC/MS m/z: 458 (M+1).

Example 13N-(8-Methyl-5,6,7,8-tetrahydro-1,6-naphthyridin-2-yl)-9-((1r,4r)-4-methylcyclohexyl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-amine

Synthesis of tert-butyl8-methyl-2-oxo-1,2,7,8-tetrahydro-1,6-naphthyridine-6(5H)-carboxylate

tert-Butyl 3-methyl-4-oxopiperidine-1-carboxylate (commerciallyavailable from Ryan Scientific, Mt. Pleasant, S.C.)(1.7 g, 7.97 mmol)and pyrrolidine (1.3 mL, 15.94 mmol) were dissolved in toluene (9 mL),and the solution was heated under reflux, with the removal of waterunder Dean-Stark conditions, for 5 hours. The solution was then cooledto room temperature and propiolamide (prepared as described in EP1813606)(1.1 g, 15.94 mmol) was added. The reaction mixture thusobtained was heated overnight under reflux. The reaction mixture wasconcentrated, and the residue was purified by flash chromatography onsilica gel eluting with 0% to 10% MeOH in DCM to give the title compound(2.1 g, 55% yield). ¹H NMR (400 MHz, CDCl₃) δ ppm 1.36 (3H, d, J=7.04Hz), 1.52 (9H, s), 3.37-3.44 (1H, m), 3.77-3.84 (1H, m), 4.12-4.17 (1H,m), 6.48 (1H, d, J=9.39 Hz), 7.21 (1H, d, J=9.39 Hz) ppm; LC/MS m/z: 265(M+1).

Synthesis of tert-butyl8-methyl-2-(((trifluoromethyl)sulfonyl)oxy)-7,8-dihydro-1,6-naphthyridine-6(5H)-carboxylate

To a solution of tert-butyl8-methyl-2-oxo-1,2,7,8-tetrahydro-1,6-naphthyridine-6(5H)-carboxylate(129 mg, 0.49 mmol) and pyridine (120 μL, 1.47 mmol) in DCM (10 mL) at0° C. was added dropwise trifluoromethanesulfonic anhydride(commercially available from Sigma-Aldrich, St. Louis, Mo.) (99 μL, 0.59mmol). The reaction mixture was stirred for 30 minutes and thenconcentrated under reduced pressure. The residue was purified by flashchromatography on silica gel eluting with 0% to 40% EtOAc in hexane togive the title compound (161 mg, 83% yield). LC/MS m/z: 397 (M+1).

Synthesis of tert-butyl8-methyl-2-((9-((1r,4r)-4-methylcyclohexyl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-yl)amino)-7,8-dihydro-1,6-naphthyridine-6(5H)-carboxylate

A glass microwave reaction vessel was charged with9-((1r,4r)-4-methylcyclohexyl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-amine(110 mg, 0.39 mmol) and(9,9-dimethyl-9H-xanthene-4,5-diyl)bis(diphenylphosphine) (45.2 mg,0.078 mmol), Pd₂(dba)₃ (35.8 mg, 0.039 mmol), tert-butyl8-methyl-2-(trifluoromethylsulfonyloxy)-7,8-dihydro-1,6-naphthyridine-6(5H)-carboxylateand 2 mL of 1,4-dioxane. The reaction mixture was heated at 100° C. for90 minutes under microwave radiation. The reaction mixture wasconcentrated, and the residue was purified by flash chromatography onsilica gel eluting with 0% to 70% solvent A (DCM:MeOH:NH₄OH, 90:9:1) inDCM to give the title compound. LCMS m/z: 528 (M+1).

Synthesis ofN-(8-methyl-5,6,7,8-tetrahydro-1,6-naphthyridin-2-yl)-94(1r,4r)-4-methylcyclohexyl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-amine(13)

A solution of tert-butyl8-methyl-249-((1r,4r)-4-methylcyclohexyl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-yl)amino)-7,8-dihydro-1,6-naphthyridine-6(5H)-carboxylatein 50% TFA in DCM (10 mL) was stirred at room temperature for 30 minutesand then concentrated and purified by preparatory LC to give the titlecompound (13) (135 mg, 81%). ¹H NMR (400 MHz, CD₃OD) δ ppm 1.10 (3H, d,J=6.46 Hz), 1.36-1.41 (2H, m), 1.56 (3H, d, J=6.65 Hz), 1.70-1.80 (1H,br. s), 2.00-2.08 (4H, m), 2.65-2.71 (2H, m), 2.73 (2H, s), 3.76-3.81(1H, m), 4.46 (2H, d, J=4.50 Hz), 4.91-4.94 (1H, m), 7.76 (1H, d, J=8.61Hz), 8.47 (1H, d, J=8.41 Hz) 8.56-8.61 (2H, m) 9.37 (1H, s) 9.54 (1H, s)ppm; LC/MS m/z: 428 (M+1).

Example 141-(8-Methyl-2-((9-((1r,4r)-4-methylcyclohexyl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-yl)amino)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)ethanone

Synthesis of1-(8-methyl-2-((9-((1r,4r)-4-methylcyclohexyl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-yl)amino)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)ethanone(14)

The title compound was prepared using chemistry similar to thatdescribed in Example 2 usingN-(8-methyl-5,6,7,8-tetrahydro-1,6-naphthyridin-2-yl)-9-((1r,4r)-4-methylcyclohexyl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-amine(13) instead of9-((1r,4r)-4-methylcyclohexyl)-N-(5,6,7,8-tetrahydro-1,6-naphthyridin-2-yl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-amine(1). ¹H NMR (400 MHz, CD₃OD) δ ppm 1.10 (3H, d, J=6.46 Hz), 1.31-1.43(2H, m), 1.48-1.57 (3H, m), 2.00-2.09 (4H, m), 2.28 (3H, s), 2.68-2.80(2H, m), 3.70-3.89 (1H, m), 3.99 (1H, td, J=13.35, 4.79 Hz), 4.91-5.01(1H, m), 7.92 (1H, br. s), 7.96-8.01 (1H, m), 8.64-8.69 (2H, m), 9.46(1H, d, J=2.93 Hz), 9.70 (1H, d, J=3.52 Hz) ppm; LC/MS m/z: 470 (M+1).

Example 152-(8-Methyl-2-((9-((1r,4r)-4-methylcyclohexyl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-yl)amino)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)ethanol

Synthesis of2-(8-methyl-2-(9-((1r,4r)-4-methylcyclohexyl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-yl)amino)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)ethanol(15)

The title compound was prepared using chemistry similar to thatdescribed in Example 3 usingN-(8-methyl-5,6,7,8-tetrahydro-1,6-naphthyridin-2-yl)-9-((1r,4r)-4-methylcyclohexyl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-amine(13) instead of9-((1r,4r)-4-methylcyclohexyl)-N-(5,6,7,8-tetrahydro-1,6-naphthyridin-2-yl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-amine(1). ¹H NMR (400 MHz, CD₃OD) δ ppm 1.10 (3H, d, J=6.65 Hz), 1.30-1.47(2H, m), 1.59 (3H, d, J=6.85 Hz), 1.69-1.83 (1H, m), 1.94-2.11 (4H, m),2.67-2.81 (2H, m), 3.39-3.58 (4H, m), 3.92-4.10 (3H, m), 4.54-4.71 (2H,m), 4.94-5.05 (1H, m), 7.80 (1H, d, J=8.80 Hz), 8.38 (1H, d, J=8.61 Hz),8.59-8.67 (2H, m), 9.42 (1H, s), 9.60 (1H, s) ppm; LC/MS m/z: 472 (M+1).

Example 162-((9-((1r,4r)-4-Methylcyclohexyl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-yl)amino)-5,6,7,8-tetrahydro-1,6-naphthyridin-8-ol

Synthesis of6-(tert-butoxycarbonyl)-2-chloro-5,6,7,8-tetrahydro-1,6-naphthyridine1-oxide

3-Chlorobenzoperoxoic acid (241 mg, 1.4 mmol) was added to a solution at0° C. of tert-butyl2-chloro-7,8-dihydro-1,6-naphthyridine-6(5H)-carboxylate (250 mg, 0.93mmol, see Example 1 procedure) in CHCl₃ (3 mL). The reaction mixturethus obtained was warmed to room temperature and stirred overnight.After concentration, the residue was purified by flash chromatography onsilica gel eluting with 0% to 80% EtOAc in hexane to give the titlecompound (150 mg, 56.6% yield). ¹H NMR (400 MHz, CDCl₃) δ ppm 1.51 (9H,s), 3.11 (2H, m), 3.77 (2H, t, J=6.16 Hz), 4.60 (2H, s), 6.99 (1H, d,J=8.41 Hz), 7.40 (1H, d, J=8.41 Hz) ppm; LC/MS m/z: 285 (M+1).

Synthesis of tert-butyl8-acetoxy-2-chloro-7,8-dihydro-1,6-naphthyridine-6(5H)-carboxylate

A solution of6-(tert-Butoxycarbonyl)-2-chloro-5,6,7,8-tetrahydro-1,6-naphthyridine1-oxide (150 mg, 0.53 mmol) in acetic anhydride (2.5 mL, 26.3 mmol), wasstirred and heated at 70° C. overnight under nitrogen atmosphere. DCM(20 mL) was added to the reaction mixture, and the resulting solutionwas washed sequentially with water, saturated aqueous NaHCO₃ solutionand brine, dried over anhydrous MgSO₄, and concentrated under vacuum.The residue was purified by flash chromatography on silica gel elutingwith 0% to 30% EtOAc in hexane to give the title compound (115 mg, 66.8%yield). ¹H NMR (400 MHz, CDCl₃) δ ppm 1.51 (9H, s), 2.13 (3H, s),3.30-3.49 (1H, d, J=12.0 Hz), 4.25-4.40 (1H, m), 4.45-4.55 (1H, d,J=12.0 Hz), 5.20-5.00 (1H, m), 5.73 (1H, br. s), 7.34 (1H, d, J=8.22Hz), 7.51 (1H, d, J=8.41 Hz) ppm; LC/MS m/z: 327 (M+1).

Synthesis of2-((9-((1r,4r)-4-methylcyclohexyl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-yl)amino)-5,6,7,8-tetrahydro-1,6-naphthyridin-8-ylacetate

The title compound was prepared using chemistry similar to thatdescribed in Example 13 using tert-butyl8-acetoxy-2-chloro-7,8-dihydro-1,6-naphthyridine-6(5H)-carboxylate inplace of tert-butyl8-methyl-2-(trifluoromethylsulfonyloxy)-7,8-dihydro-1,6-naphthyridine-6(5H)-carboxylate.LC/MS m/z: 472 (M+1).

Synthesis of(2-((9-((1r,4r)-4-methylcyclohexyl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-yl)amino)-5,6,7,8-tetrahydro-1,6-naphthyridin-8-ol(16)

Lithium hydroxide (47.6 mg, 2.0 mmol) was added to a solution of2-((9-((1r,4r)-4-methylcyclohexyl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-yl)amino)-5,6,7,8-tetrahydro-1,6-naphthyridin-8-ylacetate (106 mg, 0.226 mmol) in 10 mL of MeOH, 10 mL of THF and 5 mL ofwater. The reaction mixture was heated at 50° C. for 1 hour. Thereaction mixture was extracted with DCM, and the organic layer wasconcentrated. The residue was purified by preparative LC to give thetitle compound (16)(87 mg, 90%). ¹H NMR (400 MHz, CD₃OD) δ ppm 1.10 (3H,d, J=6.46 Hz), 1.32-1.47 (2H, m), 1.70-1.82 (1H, m), 1.99-2.11 (4H, m),2.73 (2H, qd, J=12.91, 4.30 Hz), 3.69 (2H, d, J=3.52 Hz), 4.42-4.54 (2H,m), 4.97-5.02 (2H, m), 7.89 (1H, d, J=8.61 Hz), 8.42 (1H, d, J=8.80 Hz),8.59-8.67 (2H, m), 9.43 (1H, s), 9.63 (1H, s) ppm; LC/MS m/z: 430 (M+1).

Example 176-(2-Hydroxyethyl)-2-((9-((1r,4r)-4-methylcyclohexyl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-yl)amino)-5,6,7,8-tetrahydro-1,6-naphthyridin-8-ol

Synthesis of6-(2-hydroxyethyl)-2-((9-((1r,4r)-4-methylcyclohexyl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-yl)amino)-5,6,7,8-tetrahydro-1,6-naphthyridin-8-ol(17)

The title compound was prepared using chemistry similar to thatdescribed in Example 3 using(2-((9-((1r,4r)-4-methylcyclohexyl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-yl)amino)-5,6,7,8-tetrahydro-1,6-naphthyridin-8-ol(16) in place of9-((1r,4r)-4-methylcyclohexyl)-N-(5,6,7,8-tetrahydro-1,6-naphthyridin-2-yl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-amine(1). ¹H NMR (400 MHz, CD₃OD) δ ppm 1.10 (3H, d, J=6.46 Hz), 1.32-1.46(2H, m), 1.76 (1H, dd, J=6.75, 3.81 Hz), 1.98-2.11 (4H, m), 2.67-2.80(2H, m), 3.49-3.62 (2H, m), 3.75-3.98 (2H, m), 3.98-4.10 (2H, m),4.55-4.70 (2H, m), 4.92-5.06 (2H, m), 7.86 (1H, d, J=8.80 Hz), 8.50 (1H,d, J=8.61 Hz), 8.59-8.68 (2H, m), 9.42 (1H, s), 9.59-9.64 (1H, m) ppm;LC/MS m/z: 472 (M+1).

Example 18a1((R)-8-Methyl-2-((9-((1r,4R)-4-methylcyclohexyl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-yl)amino)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)ethanoneor1((S)-8-methyl-2-((9-((1r,4S)-4-methylcyclohexyl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-yl)amino)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)ethanone

Separation of1-(8-methyl-2-((9-((1r,4r)-4-methylcyclohexyl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-yl)amino)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)ethanone

The racemic mixture obtained in Example 14 was separated on a Thar 350SFC system by sequential injections (1.2 mL; 8.0 mg/mL in MeOH; 10 mLtotal) with a 250×30 mm IA column and with 45 mL/min IPA+(20 mM NH₃) and55 g/min CO₂. The resulting fractions were concentrated on a rotaryevaporator and analyzed. The enantiomers were each obtained with >99%ee.

The first enantiomer to elute was assigned as 18a (16 mg as an off-whitesolid). ¹H NMR (500 MHz, CD₃OD:DCM-d₂ 1:1, rotamers) δ ppm 0.92-0.79(1H, m), 1.06 (3H, d, J=6.6 Hz), 1.28-1.25 (1H, m), 1.34-1.31 (2H, m),1.41-1.36 (2H, m), 1.75-1.62 (1H, m), 1.98 (4H, d, J=10.0 Hz), 2.22 (3H,d, J=4.2 Hz), 2.71-2.55 (2H, m), 3.12-2.97 (1H, m), 3.77-3.63 (1H, m),3.92-3.78 (1H, m), 4.85-4.71 (1H, m), 4.91 (1H, d, J=17.1 Hz), 7.62-7.54(1H, m), 7.98 (1H, d, J=5.4 Hz), 8.46-8.37 (2H, m), 8.92 (1H, s),9.17-9.13 (1H, m) ppm; LCMS m/z: 470.2 (M+1); HPLC purity: 99.3% (254nm).

Example 18b Stereoisomer of Example 18a. 1-((R)-8-Methyl-2-((9-((1r,4R)-4-methylcyclohexyl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-yl)amino)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)ethanoneor1-((S)-8-methyl-2-09-((1r,4S)-4-methylcyclohexyl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-yl)amino)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)ethanone

The second enantiomer to elute in the separation described in Example18a was assigned as 18b (20 mg as a light tan solid). ¹H NMR (500 MHz,CD₃OD:DCM-d₂ 1:1, rotamers) δ ppm 0.89-0.80 (m, 1H), 1.06 (3H, d, J=6.6Hz), 1.31-1.25 (2H, m), 1.43-1.35 (3H, m), 1.74-1.62 (1H, m), 2.00 (4H,d, J=10.0 Hz), 2.22 (3H, d, J=2.2 Hz), 2.71-2.58 (2H, m), 3.17-3.12 (1H,m), 3.67 (1H, dd, J=5.7, 13.6 Hz), 3.88-3.76 (1H, m), 4.92-4.80 (2H, m),7.67 (1H, d, J=8.3 Hz), 8.33-8.18 (2H, m), 8.47 (1H, d, J=5.6 Hz), 9.10(1H, s), 9.35 (1H, d, J=1.7 Hz) ppm; LCMS m/z: 470.2 (M+1); HPLC purity:98.6% (254 nm).

Example 196-Methoxy-9-((1r,4r)-4-methylcyclohexyl)-N-(5,6,7,8-tetrahydro-1,6-naphthyridin-2-yl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-amine

Synthesis of5-(5-fluoro-2-methoxypyridin-4-yl)-N4-((1r,4r)-4-methylcyclohexyl)pyrimidine-2,4-diamine

A pre-cooled (−20° C.) solution of 5-fluoro-2-methoxypyridine(commercially available from Waterstone Technology, Carmel, Ind.)(3.56g, 28.0 mmol) in THF (25 mL) was added to a 2 M lithium diisopropylamidesolution in THF (14.45 mL, 28.9 mmol) over 20 minutes at −78° C. Theyellow-brown solution thus obtained was stirred at −78° C. for 1.5hours, then zinc (II) chloride (58.7 mL, 29.4 mmol) was added. Thecooling bath was removed, and the mixture was allowed to warm to roomtemperature. To the reaction mixture was added a solution of5-iodo-N⁴((1r,4r)-4-methylcyclohexyl)pyrimidine-2,4-diamine (3 g, 9.03mmol, See Example 1 procedure) and Pd[(PPh₃)]₄ (1.044 g, 0.903 mmol) inTHF (20 mL). The resulting mixture was refluxed overnight. The reactionwas cooled to room temperature, quenched with saturated NaHCO₃(50 mL)and extracted with DCM. The organic layer was concentrated, and theresidue was purified by flash chromatography on silica gel eluting with0% to 100% EtOAc in hexane to give the title compound as a yellow solid(1.5 g, 50% yield). ¹H NMR (400 MHz, DMSO-d₆) δ ppm 0.86 (3H, d, J=8.0Hz), 0.89-1.00 (2H, m), 1.16-1.32 (3H, m), 1.65 (2H, d, J=12.0 Hz), 1.76(2H, d, J=12.0 Hz), 3.84 (3H, s), 3.92-4.02 (1H, m), 5.96 (1H, d, J=12.0Hz), 6.20 (2H, s), 6.74 (1H, d, J=8.0 Hz), 7.57 (1H, s), 8.10 (1H, s)ppm; LCMS m/z: 332.2 (M+1).

Synthesis of6-methoxy-9-((1r,4r)-4-methylcyclohexyl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-amine.

Sodium hydride (60% dispersion in mineral oil) (1.09 g, 27.2 mmol) wasadded slowly to a solution of5-(5-fluoro-2-methoxypyridin-4-yl)-N⁴-((1r,4r)-4-methylcyclohexyl)pyrimidine-2,4-diamine(3.0 g, 9.05 mmol) in DMF (60 mL) at 0° C. After addition, the mixturewas stirred at room temperature for 20 minutes under N₂ atmosphere andthen at 105° C. overnight. The reaction was cooled to room temperatureand quenched with saturated NH₄Cl at 0° C. while stirring. The mixturewas extracted with EtOAc and washed with water and then with brine, anddried over Na₂SO₄. The solvent was removed, and the crude product waspurified by flash chromatography on silica gel eluting 0% to 50% EtOAcin hexane to give the title compound as an off-white solid (1.5 g, 50%yield). ¹H NMR (400 MHz, DMSO-d₆) δ ppm 0.96 (3H, d, J=8.0 Hz),1.05-1.24 (2H, m), 1.55-1.65 (1H, m), 1.74 (2H, d, J=8.0 Hz), 1.85 (2H,d, J=8.0 Hz), 2.30-2.45 (2H, m), 3.87 (3H, s), 4.64-4.74 (1H, m), 6.93(2H, s), 7.31 (1H, s), 8.52 (1H, s), 8.94 (1H, s) ppm; LCMS m/z: 312.3(M+1).

Synthesis of6-methoxy-9-((1r,4r)-4-methylcyclohexyl)-N-(5,6,7,8-tetrahydro-1,6-naphthyridin-2-yl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-amine(19)

The title compound was prepared using chemistry similar to thatdescribed in Example 1 using6-methoxy-9-((1r,4r)-4-methylcyclohexyl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-aminein place of9-((1r,4r)-4-methylcyclohexyl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-amine.¹H NMR (500 MHz, CD₃OD) δ ppm 1.05 (3H, d, J=6.6 Hz), 1.25-1.48 (2H, m),1.65-1.75 (1H, m), 1.95-2.05 (4H, m), 2.51-2.74 (2H, m), 3.42 (2H, d,J=10.0 Hz), 3.70 (2H, d, J=10.0 Hz), 4.03 (3H, s), 4.45 (2H, s),4.82-4.90 (1H, m), 7.30-7.45 (1H, m), 7.62 (1H, s), 7.84 (1H, d, J=10.0Hz), 8.73 (1H, s), 9.35 (1H, s) ppm; LCMS m/z: 444.3 (M+1).

Example 202-Hydroxy-1-(2-((6-methoxy-9-((1r,4r)-4-methylcyclohexyl)-9H-pyrido[4′,3′:4,5]-pyrrolo[2,3-d]pyrimidin-2-yl)amino)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)ethanone

Synthesis of2-hydroxy-1-(2-((6-methoxy-9-((1r,4r)-4-methylcyclohexyl)-9H-pyrido[4′,3′:4,5]-pyrrolo[2,3-d]pyrimidin-2-yl)amino)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)ethanone(20)

The title compound was prepared using chemistry similar to thatdescribed in Example 5 using6-methoxy-9-((1r,4r)-4-methylcyclohexyl)-N-(5,6,7,8-tetrahydro-1,6-naphthyridin-2-yl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-amine(19) in place of9-((1r,4r)-4-methylcyclohexyl)-N-(5,6,7,8-tetrahydro-1,6-naphthyridin-2-yl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-amine(1). ¹H NMR (500 MHz, CD₃OD) δ ppm 1.04 (3H, d, J=6.6 Hz), 1.25-1.33(2H, m), 1.65-1.75 (1H, m), 1.95-2.05 (4H, m), 2.51-2.62 (2H, m),3.14-3.26 (2H, m), 3.84-3.99 (2H, m), 4.02 (3H, s), 4.37 (2H, br. s.),4.74 (2H, br. s.), 4.82-4.91 (1H, m), 7.28 (1H, d, J=10.0 Hz), 7.59 (1H,s), 7.89-7.94 (1H, m), 8.70 (1H, s), 9.34 (1H, s) ppm; LCMS m/z: 502.2(M+1).

Example 219-Cyclopentyl-N-(5,6,7,8-tetrahydro-1,6-naphthyridin-2-yl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-amine

Synthesis of tert-butyl2-((9-cyclopentyl-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-yl)amino)-7,8-dihydro-1,6-naphthyridine-6(5H)-carboxylate

To a solution of9-cyclopentyl-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-amine(prepared as described in WO 2009/085185) (152 mg, 0.6 mmol) in dioxane(6 mL) were added tert-butyl2-chloro-7,8-dihydro-1,6-naphthyridine-6(5H)-carboxylate (177 mg, 0.66mmol), tris(dibenzylideneacetone)dipalladium (0) (28 mg, 0.030 mmol),4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (52 mg, 0.090 mmol), andsodium t-butoxide (86 mg, 0.9 mmol). The reaction mixture thus obtainedwas heated at 150° C. under microwave irradiation for 1 hour. Thereaction mixture was diluted with DCM, washed with brine, and thendried. The solvent was evaporated and the residue was purified by flashchromatography on silica gel eluting with 10% to 50% solvent A(DCM/MeOH/NH₄OH, 100:10:1) in DCM to give the title compound as a lightyellow solid (211 mg, 72% yield). ¹H NMR (500 MHz, CDCl₃) δ 1.52 (9H,s), 1.88-1.90 (2H, m), 2.14-2.21 (4H, m), 2.45-2.47 (2H, m), 2.92-2.94(2H, m), 3.76-3.79 (2H, m), 4.59 (2H, m), 5.37 (1H, m), 7.49 (1H, d,J=10.0 Hz), 7.85 (1H, d, J=5.0 Hz), 8.14 (1H, br. s), 8.37 (1H, d,J=10.0 Hz), 8.52 (1H, d, J=5.0 Hz), 8.92 (1H, s), 9.11 (1H, s) ppm; LCMSm/z: 486 (M+1).

Synthesis of9-cyclopentyl-N-(5,6,7,8-tetrahydro-1,6-naphthyridin-2-yl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-amine(21).

A solution of tert-butyl2-((9-cyclopentyl-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-yl)amino)-7,8-dihydro-1,6-naphthyridine-6(5H)-carboxylate(217 mg, 0.477 mmol) in TFA/DCM (1:1, 2 mL) was stirred at roomtemperature for 30 minutes and then concentrated. The residue waspurified by flash chromatography on silica gel eluting with 25% to 75%solvent A (DCM:MeOH:NH₄OH, 100:10:1) in DCM to give the title compoundas a yellow solid (148 mg, 86% yield). ¹H NMR (500 MHz, CDCl₃) δ1.87-1.90 (2H, m), 2.14-2.21 (4H, m), 2.43-2.47 (2H, m), 2.89 (2H, t,J=5.0 Hz), 3.26 (2H, t, J=5.0 Hz), 4.02 (2H, s), 5.36 (1H, m), 7.41 (1H,d, J=10.0 Hz), 7.85 (1H, d, J=5.0 Hz), 8.19 (1H, br. s), 8.31 (1H, d,J=10.0 Hz), 8.52 (1H, d, J=5.0 Hz), 8.91 (1H, s), 9.11 (1H, s) ppm; LCMSm/z: 386 (M+1).

Example 221-(2-((9-Cyclopentyl-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-yl)amino)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)-2-hydroxyethanone

Synthesis of1-(2-((9-cyclopentyl-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-yl)amino)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)-2-hydroxyethanone(22)

To a solution of9-cyclopentyl-N-(5,6,7,8-tetrahydro-1,6-naphthyridin-2-yl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-amine(21) (145 mg, 0.367 mmol) in DMF (10 mL) were added 2-hydroxyacetic acid(34.3 mg, 0.451 mmol), EDC (191.7 mg, 0.451 mmol),N-hydroxybenzotriazole (61 mg, 0.451 mmol), andN,N-diisopropylethylamine (157 uL, 0.903 mmol). The mixture thusobtained was stirred at room temperature for 2 hours. The reactionmixture was concentrated, and the residue was purified by flashchromatography on silica gel eluting with 20% to 75% solvent A(DCM:MeOH:NH₄OH, 100:10:1) to give the title compound as an off-whitesolid (117 mg, 70% yield). ¹H NMR (500 MHz, DMSO-d₆) δ 1.77-1.80 (2H,m), 2.07-2.10 (4H, m), 2.42-2.44 (2H, m), 2.82-2.91 (2H, m), 3.66-3.82(2H, m), 4.18-4.20 (2H, m), 4.57-4.66 (2H, m), 5.36 (1H, m), 7.68 (1H,d, J=10.0 Hz), 8.06 (1H, d, J=5.0 Hz), 8.19 (1H, d, J=10.0 Hz), 8.48(1H, d, J=5.0 Hz), 9.03 (1H, s), 9.31 (1H, s) 9.98 (1H, br. s) ppm; LCMSm/z: 444 (M+1).

Example 239-((1r,4r)-4-Methylcyclohexyl)-N-(5,6,7,8-tetrahydro-1,7-naphthyridin-2-yl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-amine

Synthesis of tert-butyl2-chloro-5,6-dihydro-1,7-naphthyridine-7(8H)-carboxylate

The title compound was prepared using chemistry similar to thatdescribed in Example 1 using2-chloro-5,6,7,8-tetrahydro-1,7-naphthyridine hydrochloride(commercially obtained from Anichem, North Brunswick, N.J.) in place of2-chloro-5,6,7,8-tetrahydro-1,6-naphthyridine hydrochloride. ¹H NMR (500MHz, CD₂Cl₂) δ 1.47 (9H, s), 2.79 (2H, t, J=5.6 Hz), 3.64 (2H, t, J=5.6Hz), 4.58 (2H, s), 7.13 (1H, d, J=8.0 Hz), 7.42 (1H, d, J=8.0 Hz) ppm;LCMS m/z: 269 (M+1).

Synthesis of tert-butyl2-((9-((1r,4r)-4-methylcyclohexyl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-yl)amino)-5,6-dihydro-1,7-naphthyridine-7(8H)-carboxylate

The title compound was prepared using chemistry similar to thatdescribed in Example 1 using tert-butyl2-chloro-5,6-dihydro-1,7-naphthyridine-7(8H)-carboxylate in place oftert-butyl 2-chloro-7,8-dihydro-1,6-naphthyridine-6(5H)-carboxylate. ¹HNMR (500 MHz, CDCl₃) δ 1.07 (3H, d, J=6.4 Hz), 1.23-1.33 (2H, m), 1.52(9H, s), 1.64-1.68 (1H, m), 1.99 (4H, t, J=11.0 Hz), 2.60 (2H, dq,J=3.3, 12.3 Hz), 2.85 (2H, br. s), 3.72 (2H, br. s), 4.60 (2H, s), 4.73(1H, t, J=12.3 Hz), 7.53 (1H, d, J=8.3 Hz), 7.87 (1H, d, J=5.4 Hz), 8.10(1H, br. s), 8.37 (1H, d, J=8.3 Hz), 8.51 (1H, d, J=5.4 Hz), 8.97 (1H,s), 9.12 (1H, s) ppm; LCMS m/z: 514 (M+1).

Synthesis of9-((1r,4r)-4-methylcyclohexyl)-N-(5,6,7,8-tetrahydro-1,7-naphthyridin-2-yl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-aminedihydrochloride

To a solution of tert-butyl2-((9-((1r,4r)-4-methylcyclohexyl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-yl)amino)-5,6-dihydro-1,7-naphthyridine-7(8H)-carboxylate(320 mg, 0.623 mmol) in DCM (5 mL) and MeOH (5 mL) was added 4.0 M HClin dioxane (10 mL) and a slurry formed within minutes. The mixture wasstirred overnight at room temperature, concentrated and then reslurriedwith ether. The slurry was filtered with a Buchner funnel providing9-((1r,4r)-4-methylcyclohexyl)-N-(5,6,7,8-tetrahydro-1,7-naphthyridin-2-yl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-aminedihydrochloride (176 mg) as a tan solid. ¹H NMR (500 MHz, CD₃OD) δ 1.09(3H, d, J=6.4 Hz), 1.32-1.45 (2H, m), 1.77 (1H, br. s), 2.06 (4H, t,J=15.5 Hz), 2.64-2.79 (2H, m), 3.21 (2H, t, J=6.2 Hz), 3.63 (2H, t,J=6.2 Hz), 4.61 (2H, s), 4.93-5.03 (1H, m), 7.88 (1H, br. s), 7.91-7.96(1H, m), 8.74 (1H, d, J=6.1 Hz), 8.81 (1H, d, J=6.1 Hz), 9.55 (1H, s),9.80 (1H, s) ppm; LCMS m/z: 414 (M+1).

Example 249-((1r,4r)-4-Methylcyclohexyl)-N-(5,6,7,8-tetrahydro-2,7-naphthyridin-3-yl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-amine

Synthesis of tert-butyl6-chloro-3,4-dihydro-2,7-naphthyridine-2(1H)-carboxylate

6-Chloro-1,2,3,4-tetrahydro-2,7-naphthyridine hydrochloride(commercially obtained from Anichem, North Brunswick, N.J.) (2.0 g, 9.75mmol) and TEA (2.72 mL, 19.50 mmol) were mixed in DCM (19.50 mL). Boc₂O(2.34 g, 10.7 mmol) was added. The mixture was then stirred for 2 hoursat room temperature. TLC indicated good consumption of the startingmaterial and the formation of one major spot. There was also a lesspolar spot that was more faint. The reaction mixture was partitionedwith DCM and water. The organic layer was dried with sodium sulfate,filtered, and concentrated. The residue was purified by flash columnchromatography on silica gel eluting with 0% to 40% EtOAc in hexane toprovide tert-butyl6-chloro-3,4-dihydro-2,7-naphthyridine-2(1H)-carboxylate as a clearcolorless oil (1.53 g). ¹H NMR (500 MHz, CD₂CCl₂) δ 1.46 (9H, s), 2.80(2H, t, J=6.0 Hz), 3.62 (2H, t, J=6.0 Hz), 4.55 (2H, s), 7.12 (1H, s),8.13 (1H, s) ppm; LCMS m/z: 269 (M+1).

Synthesis of tert-butyl6-((9-((1r,4r)-4-methylcyclohexyl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-yl)amino)-3,4-dihydro-2,7-naphthyridine-2(1H)-carboxylate

The title compound was prepared using chemistry similar to thatdescribed in Example 1 using tert-butyl6-chloro-3,4-dihydro-2,7-naphthyridine-2(1H)-carboxylate in place oftert-butyl 2-chloro-7,8-dihydro-1,6-naphthyridine-6(5H)-carboxylate. ¹HNMR (500 MHz, CDCl₃) δ 1.07 (3H, d, J=6.4 Hz), 1.23-1.35 (2H, m), 1.53(9H, s), 1.62-1.73 (1H, m), 2.00 (2H, br. s), 2.48-2.67 (2H, m),2.81-3.01 (2H, m), 3.72 (2H, br. s), 4.61 (2H, s), 4.78 (1H, t, J=12.5Hz), 7.89 (1H, d, J=5.4 Hz), 8.10 (1H, br. s), 8.12 (1H, br. s), 8.36(1H, br. s), 8.52 (1H, d, J=5.4 Hz), 8.98 (1H, s), 9.12 (1H, s) ppm;LCMS m/z: 514 (M+1).

Synthesis of9-((1r,4r)-4-methylcyclohexyl)-N-(5,6,7,8-tetrahydro-2,7-naphthyridin-3-yl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-aminedihydrochloride

To a slurry of tert-butyl6-((9-((1r,4r)-4-methylcyclohexyl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-yl)amino)-3,4-dihydro-2,7-naphthyridine-2(1H)-carboxylate(446 mg, 0.868 mmol) in MeOH (4.3 mL) and DCM (4.3 mL) was added 10 mLof 4.0 M HCl in dioxane. The resulting clear solution was then stirredovernight at room temperature. Solid precipitated from the reactionmixture during the course of the reaction. The mixture was concentrated,rinsed with ether and filtered to yield9-((1r,40-4-methylcyclohexyl)-N-(5,6,7,8-tetrahydro-2,7-naphthyridin-3-yl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-aminedihydrochloride (461 mg, 0.948 mmol). ¹H NMR (500 MHz, CD₃OD) δ 1.07(3H, d, J=6.6 Hz), 1.29-1.42 (2H, m), 1.69-1.81 (1H, m), 1.98-2.09 (4H,m), 2.64-2.76 (2H, m), 3.39 (2H, t, J=6.4 Hz), 3.64 (2H, t, J=6.4 Hz),4.54 (2H, s), 4.95-5.07 (1H, m), 7.68 (1H, br. s), 8.48 (1H, s), 8.71(1H, d, J=6.1 Hz), 8.83 (1H, d, J=6.1 Hz), 9.55 (1H, s), 9.78 (1H, s)ppm; LCMS m/z: 414 (M+1).

Example 251-(2-((9-Cycloheptyl-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-yl)amino)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)-2-hydroxyethanone

Synthesis of1-(2-((9-cycloheptyl-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-yl)amino)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)-2-hydroxyethanone(25)

The title compound can be prepared using chemistry similar to thatdescribed in Examples 1 and 5 using cycloheptylamine (commerciallyavailable from Sigma-Aldrich, St. Louis, Mo.) in place oftrans-4-methylcyclohexylamine hydrochloride.

Example 261-(2-β9-Cyclopentyl-8-fluoro-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-yl)amino)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)-2-hydroxyethanone

Synthesis of1-(2-((9-cyclopentyl-8-fluoro-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-yl)amino)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)-2-hydroxyethanone(26)

The title compound can be prepared using chemistry similar to thatdescribed in Examples 21 and 22 using9-cyclopentyl-8-fluoro-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-amine(prepared as described in WO 2009/085185) in place of9-cyclopentyl-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-amine.

Example 272-Hydroxy-1-(2-((9-(tetrahydro-2H-pyran-4-yl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-yl)amino)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)ethanone

Synthesis of2-hydroxy-1-(2-((9-(tetrahydro-2H-pyran-4-yl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-yl)amino)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)ethanone(27)

The title compound can be prepared using chemistry similar to thatdescribed in Examples 1 and 5 using9-(tetrahydro-2H-pyran-4-yl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-amine(prepared as described in WO 2009/085185) in place of9-((1r,4r)-4-methylcyclohexyl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-amine.

Example 28 1-(2-((9-((1R,2R,4S)-Bicyclo[2.2.1]heptan-2-yl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-yl)amino)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)-2-hydroxyethanone

Synthesis of 1-(2-((9-((1R,2R,4S)-bicyclo[2.2.1]heptan-2-yl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-yl)amino)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)-2-hydroxyethanone(28)

The title compound can be prepared using chemistry similar to thatdescribed in Examples 1 and 5 using9-((1R,2R,4S)-bicyclo[2.2.1]heptan-2-yl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-amine(prepared as described in WO 2009/085185) in place of9-((1r,4r)-4-methylcyclohexyl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-amine.

Example 291-(2-((9-((1s,3s)-Adamantan-1-yl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-yl)amino)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)-2-hydroxyethanone

Synthesis of1-(2-((9-((1s,3s)-adamantan-1-yl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-yl)amino)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)-2-hydroxyethanone(29)

The title compound can be prepared using chemistry similar to thatdescribed in Examples 1 and 5 using9-((1s,3s)-adamantan-1-yl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-amine(prepared as described in WO 2009/085185) in place of9-((1r,4r)-4-methylcyclohexyl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-amine.

Example 302-Hydroxy-1-(2-((9-((1r,4r)-4-methylcyclohexyl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-yl)amino)-5,6-dihydro-1,7-naphthyridin-7(8H)-yl)ethanone

Synthesis of2-hydroxy-1-(2-((9-((1r,4r)-4-methylcyclohexyl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-yl)amino)-5,6-dihydro-1,7-naphthyridin-7(8H)-yl)ethanone

The title compound was prepared using chemistry similar to thatdescribed in Example 5 using9-((1r,4r)-4-methylcyclohexyl)-N-(5,6,7,8-tetrahydro-1,7-naphthyridin-2-yl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-aminedihydrochloride (Example 23) in place of9-((1r,4r)-4-methylcyclohexyl)-N-(5,6,7,8-tetrahydro-1,6-naphthyridin-2-yl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-amine.¹H NMR (500 MHz, CDCl₃) δ 1.08 (3H, d, J=6.4 Hz), 1.21-1.35 (2H, m),1.99 (4H, t, J=10.5 Hz), 2.60 (2H, dq, J=3.5, 12.8 Hz), 2.91 (2H, q,J=5.9 Hz), 3.58 (1H, t, J=5.9 Hz), 3.68 (1H, t, J=4.3 Hz), 3.97 (1H, t,J=5.9 Hz), 4.10 (2H, m), 4.44 (1H, s), 4.75 (1H, m), 4.82 (1H, s), 7.55(1H, d, J=8.6 Hz), 7.88 (1H, d, J=5.4 Hz), 8.07 (1H, s), 8.43 (1H, d,J=8.6 Hz), 8.53 (1H, d, J=5.4 Hz), 8.99 (1H, s), 9.11 (1H, s) ppm; LCMSm/z: 472 (M+1).

Example 312-Hydroxy-1-(6-((9-((1r,4r)-4-methylcyclohexyl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-yl)amino)-3,4-dihydro-2,7-naphthyridin-2(1H)-yl)ethanone

Synthesis of2-hydroxy-1-(6-((9-((1r,4r)-4-methylcyclohexyl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-yl)amino)-3,4-dihydro-2,7-naphthyridin-2(1H)-yl)ethanone

The title compound was prepared using chemistry similar to thatdescribed in Example 5 using9-((1r,4r)-4-methylcyclohexyl)-N-(5,6,7,8-tetrahydro-2,7-naphthyridin-3-yl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-aminedihydrochloride (Example 24) in place of9-((1r,4r)-4-methylcyclohexyl)-N-(5,6,7,8-tetrahydro-1,6-naphthyridin-2-yl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-amine.¹H NMR (500 MHz, CDCl₃) δ 1.07 (3H, d, J=6.4 Hz), 1.25-1.36 (3H, m),2.01 (4H, d, J=10.5 Hz), 2.51-2.64 (2H, m), 3.00 (2H, q, J=5.7 Hz), 3.59(1H, t, J=6.0 Hz), 3.61-3.67 (1H, m), 3.97 (1H, t, J=6.0 Hz), 4.31 (2H,t, J=3.4 Hz), 4.49 (1H, s), 4.71-4.85 (2H, m), 7.09 (1H, d, J=5.1 Hz),8.14 (1H, s), 8.16 (1H, s), 8.42 (1H, s), 8.52 (1H, d, J=5.1 Hz), 9.00(1H, s), 9.13 (1H, s) ppm; LCMS m/z: 472 (M+1).

Example 32(S)-2-Hydroxy-1-(2-((9-((1r,4r)-4-methylcyclohexyl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-yl)amino)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)propan-1-one

Synthesis of(S)-2-hydroxy-1-(2-((9-((1r,4r)-4-methylcyclohexyl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-yl)amino)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)propan-1-one

To a solution of9-((1r,4r)-4-methylcyclohexyl)-N-(5,6,7,8-tetrahydro-1,6-naphthyridin-2-yl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-amine(Example 1) (83 mg, 0.20 mmol) in chloroform (10 mL) were added L-lacticacid (commercially obtained from Sigma-Aldrich, St. Louis, Mo.) (15 uL,0.20 mmol), EDC (46 mg, 0.24 mmol), N-hydroxybenzotriazole (32 mg, 0.24mmol), and N,N-diisopropylethylamine (84 uL, 0.48 mmol). The mixturethus obtained was stirred at room temperature and HPLC-MS analysisindicated that the reaction was complete after 4 hours. The reactionmixture was diluted with chloroform and washed with water and brine,then the organic layer was dried over MgSO₄ and concentrated underreduced pressure. The residue was purified by flash columnchromatography on silica gel eluting with 10% to 30% solvent A(DCM:MeOH:NH₄OH, 90:9:1) in DCM to provide the title compound as a lightyellow solid (67 mg, 69% yield). ¹H NMR (500 MHz, CDCl₃) δ 1.06 (3H, d,J=5.0 Hz), 1.24-1.33 (2H, m), 1.38-1.44 (3H, m), 1.63-1.68 (1H, m),1.96-2.02 (4H, m), 2.56-2.64 (2H, m), 2.98-3.03 (2H, m), 3.78 (1H, t,J=5.0 Hz), 3.90-3.94 (1H, m), 4.12-4.15 (1H, m), 4.56-4.63 (2H, m),4.73-4.77 (1H, m), 4.81 (1H, br. s), 7.50 (1H, d, J=10 Hz), 7.54 (1H, d,J=10 Hz), 8.30 (1H, s), 8.43 (1H, d, J=10 Hz), 8.47 (1H, d, J=10 Hz),9.14 (1H, s), 9.16 (1H, s) ppm; LCMS m/z: 486 (M+1).

Example 33 (R)-2-Hydroxy-1-(2-((9-((1r,4r)-4-methylcyclohexyl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-yl)amino)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)propan-1-one

Synthesis of(R)-2-hydroxy-1-(2-((9-((1r,4r)-4-methylcyclohexyl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-yl)amino)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)propan-1-one.The title compound was prepared using chemistry similar to thatdescribed in Example 32 using D-lactic acid (commercially obtained fromSigma-Aldrich, St. Louis, Mo.) in place of L-lactic acid. ¹H NMR (500MHz, CDCl₃) δ 1.06 (3H, d, J=5.0 Hz), 1.23-1.31 (2H, m), 1.38-1.44 (3H,m), 1.63-1.67 (1H, m), 1.96-2.02 (4H, m), 2.56-2.64 (2H, m), 2.99-3.05(2H, m), 3.78 (1H, t, J=5.0 Hz), 3.90-3.93 (1H, m), 4.12-4.15 (1H, m),4.58-4.63 (2H, m), 4.72-4.77 (1H, m), 4.81 (1H, d, J=5.0 Hz), 7.50 (1H,d, J=10 Hz), 7.54 (1H, d, J=10 Hz), 7.84 (1H, s), 8.44 (1H, d, J=10 Hz),8.47 (1H, d, J=10 Hz), 9.19 (1H, s), 9.23 (1H, s) ppm; LCMS m/z: 486(M+1). Example 342-(Dimethylamino)-1-(2-((9-((1r,4r)-4-methylcyclohexyl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-yl)amino)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)ethanone

Synthesis of2-(dimethylamino)-1-(2-((9-((1r,4r)-4-methylcyclohexyl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-yl)amino)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)ethanone.The title compound was prepared using chemistry similar to thatdescribed in Example 32 using 2-(dimethylamino)acetic acid (commerciallyobtained from Sigma-Aldrich, St. Louis, Mo.) in place of L-lactic acid.¹H NMR (500 MHz, CDCl₃) δ 1.07 (3H, d, J=5.0 Hz), 1.24-1.32 (2H, m),1.64-1.67 (1H, m), 1.96-2.02 (4H, m), 2.32 (6H, s), 2.57-2.64 (2H, m),2.97 (1H, t, J=5.0 Hz), 3.02 (1H, t, J=5.0 Hz), 3.25 (2H, br. s),3.93-3.98 (2H, m), 4.70-4.76 (3H, m), 7.50 (1H, d, J=10 Hz), 7.84 (1H,d, J=5.0 Hz), 8.44 (1H, d, J=10 Hz), 8.51 (1H, d, J=5.0 Hz), 8.97 (1H,s), 9.18 (1H, s) ppm; LCMS m/z: 499 (M+1). Example 35 tert-Butyl2-((9-((1r,4r)-4-methylcyclohexyl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-yl)amino)-5,6-dihydro-1,7-naphthyridine-7(8H)-carboxylate

Synthesis of tert-butyl2-((9-((1r,4r)-4-methylcyclohexyl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-yl)amino)-5,6-dihydro-1,7-naphthyridine-7(8H)-carboxylate.The title compound was prepared as described in Example 23. ¹H NMR (500MHz, CDCl₃) δ 1.07 (3H, d, J=6.4 Hz), 1.23-1.33 (2H, m), 1.52 (9H, s),1.64-1.68 (1H, m), 1.99 (4H, t, J=11.0 Hz), 2.60 (2H, dq, J=3.3, 12.3Hz), 2.85 (2H, br. s), 3.72 (2H, br. s), 4.60 (2H, s), 4.73 (1H, t,J=12.3 Hz), 7.53 (1H, d, J=8.3 Hz), 7.87 (1H, d, J=5.4 Hz), 8.10 (1H,br. s), 8.37 (1H, d, J=8.3 Hz), 8.51 (1H, d, J=5.4 Hz), 8.97 (1H, s),9.12 (1H, s) ppm; LCMS m/z: 514 (M+1). Example 36 tert-Butyl6-((9-((1r,4r)-4-methylcyclohexyl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-yl)amino)-3,4-dihydro-2,7-naphthyridine-2(1H)-carboxylate

Synthesis of tert-butyl6-((9-((1r,4r)-4-methylcyclohexyl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-yl)amino)-3,4-dihydro-2,7-naphthyridine-2(1H)-carboxylate.The title compound was prepared as described in Example 24. ¹H NMR (500MHz, CDCl₃) δ 1.07 (3H, d, J=6.4 Hz), 1.23-1.35 (2H, m), 1.53 (9H, s),1.62-1.73 (1H, m), 2.00 (2H, br. s), 2.48-2.67 (2H, m), 2.81-3.01 (2H,m), 3.72 (2H, br. s), 4.61 (2H, s), 4.78 (1H, t, J=12.5 Hz), 7.89 (1H,d, J=5.4 Hz), 8.10 (1H, br. s), 8.12 (1H, br. s), 8.36 (1H, br. s), 8.52(1H, d, J=5.4 Hz), 8.98 (1H, s), 9.12 (1H, s) ppm; LCMS m/z: 514 (M+1).Example 372-(Diethylamino)-1-(2-((9-((1r,4r)-4-methylcyclohexyl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-yl)amino)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)ethanone

Synthesis of2-(diethylamino)-1-(2-((9-((1r,4r)-4-methylcyclohexyl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-2-yl)amino)-7,8-dihydro-1,6-naphthyridin-6(5H)-yl)ethanone.The title compound was prepared using chemistry similar to thatdescribed in Example 32 using 2-(diethylamino)acetic acid (commerciallyobtained from Matrix Scientific, Columbia, S.C.) in place of L-lacticacid. ¹H NMR (500 MHz, CDCl₃) δ ppm 1.02-1.09 (9H, m), 1.24-1.33 (2H,m), 1.63-1.69 (1H, m), 1.96-2.02 (4H, m), 2.57-2.65 (6H, m), 2.98 (2H,t, J=5.0 Hz), 3.39 (2H, br. s), 3.97 (2H, t, J=5.0 Hz), 4.71-4.77 (3H,m), 7.49 (1H, d, J=10 Hz), 7.84 (1H, d, J=5.0 Hz), 8.24 (1H, s), 8.42(1H, d, J=10 Hz), 8.51 (1H, d, J=5.0 Hz), 8.97 (1H, s), 9.13 (1H, s);LCMS m/z: 527 (M+1).

CDK4, FLT3, and MOLM13 Assays

The CDK4 and CDK1 inhibitory activity of the CDK4/6-FLT3 inhibitors wasdetermined with a filtration kinase assay. The compounds, kinase andsubstrate diluted in the kinase buffer (20 mM Tris, pH7.4, 50 mM NaCl, 1mM DTT, 0.1% BSA) were sequentially added to a 96-well Multiscreen HTSfiltration plate (Millipore). The final 100 μL reaction mixture in eachwell contained 0.3 μg of CDK4/Cyclin D1 or CDK1/Cyclin B (Cell SignalingTechnology), 1 μg of Rb fragment (aa773-928, Millipore) for the CDK4assay or 5 μg of histone H1 for the CDK1 assay and 1 μCi of [³³P]-ATP.The mixture was incubated at room temperature for 1 hour. The proteinsin the reaction were then precipitated and washed with cold TCA solutionusing an aspiration/filtration vacuum system. The plates were dried atroom temperature, and the retained radioactivity was measured byscintillation counting.

The FLT3 inhibitory activity of the CDK4/6-FLT3 inhibitors wasdetermined with a HTRF kinase assay. The FLT3 enzyme (GST-FLT3 fusion)was purchased from Carna Biosciences. An ULight-labeled syntheticpeptide derived from human Janus kinase 1 (aa1015-1027, ULight-JAK1,PerkinElmer) was utilized as the phosphoacceptor substrate. The assaywas conducted in a 384-well white OptiPlate (PerkinElmer). The 20 μLreaction mixture contained 50 nM ULight-JAK1, 116 μM ATP, 0.0385 ng/μLFLT3 and dilutions of test compounds in the kinase buffer (50 mM Hepes,pH 7.6, 1 mM EGTA, 10 mM MgCl₂, 2 mM DTT, and 0.005% Tween 20). Thereaction was allowed to proceed for 1 hour at room temperature and wasstopped by adding 20 μl., of 10 mM EDTA, 2 nM LANCE® Eu-W1024anti-phospho-tyrosine antibody in LANCE® detection buffer (PerkinElmer).The plates were incubated at room temperature for 2 hours after additionof detection reagents and then read on an Envision multimode reader(PerkinElmer).

The cell proliferation inhibition potency of the CDK4/6-FLT3 inhibitorswas determined by using a [¹⁴C]-thymidine incorporation assay.Exponentially growing cells (MOLM-13, Colo-205, etc.) were seeded in a96-well Cytostar T plate (GE Healthcare Biosciences) at a density of5×10³ cells/well and incubated overnight. Serially diluted compounds and0.1 μCi of [¹⁴C]-thymidine (GE Healthcare Biosciences) were added toeach well on the following day. After 72 hour incubation, isotopeincorporation was determined with a β plate counter (Wallac). MOLM13 isa human AML tumor cell line expressing FLT3, FLT3^(ITD) and wild typeRb.

IC₅₀ values of the compounds in the above assays were determined bynon-linear regression analysis using Prism (GraphPad Software).

The following table includes IC₅₀ values obtained using the proceduresset forth above for the Example compounds described herein.

Table of FLT3, CDK4, and MOLM13 Data For Example Compounds FLT3 CDK4MOLM13 IC₅₀ IC₅₀ IC₅₀ Example Structure^(a) (μM)^(b) (μM)^(b) (μM)^(b) 1

0.0013 0.0021 0.0152  2

0.0032 0.0045 0.0225  3

0.0018 0.0026 0.0148  4

0.002  0.0046 0.0211  5

0.0015 0.0024 0.019   6

0.0033 0.12  0.046   7

0.0214 0.0524 0.206   8

0.0361 0.0024 0.0091  9

0.0034 0.0728 0.0649 10

0.0017 0.0105 0.0332 11

0.0737 0.257  >3    12

0.0027 0.0184 0.0108 13

0.0018 0.0313 0.0545 14

0.0032 0.0044 0.0106 15

0.0031 0.0203 0.0295 16

0.0014 0.0165 0.0239 17

0.0029 0.0192 0.0219  18a

0.0016 0.0188 0.0242  18b The enantiomer of 18a 0.0023 0.463  0.02  19

0.033  1.79  0.204  20

0.144  0.847  0.208  21

ND ND ND 22

 0.00065 0.036  0.0052 23

ND 0.0086 0.0015 24

ND 0.0047 0.0035 25

ND ND ND 26

ND ND ND 27

ND ND ND 28

ND ND ND 29

ND ND ND 30

ND 0.0063 0.003  31

ND 0.035  0.0067 32

ND 0.012  0.0018 33

ND 0.041  0.0031 34

ND 0.003  0.0003 35

ND 0.026  0.48  36

ND 0.35  — 37

ND 0.003  0.0014 ^(a)The

 symbol indicates that a mixture of the R and S isomers is present withrespect to the stereocenter shown. ^(b)ND means not determined

Xenograft Studies

Tumor Growth Inhibition in AML MOLM 13 Xenograft Tumors

The anti-tumor activity of Example 5 against subcutaneous MOLM13xenograft tumors was demonstrated after treatment with increasing dosesof Example 5. The AML cell line, MOLM13, (obtained from American TypeCulture Collection, Manassas, Va., USA) was cultured in RPMI-1640 cellgrowth medium containing 10% fetal bovine serum (commercially obtainedfrom Invitrogen, Carlsbad, Calif. USA). This cell line expresses CDK4/6,one copy of wild type FLT3 kinase and one copy of FLT3-ITD, whichresults in constitutive activation of FLT3 activity. Therefore, theactivity of Example 5 against both FLT3 and CDK4/6 can be tested in thistumor cell. Tumor cells (7.5 million) were injected subcutaneously ontothe right flank of CrTac:NCR-FoxnF nude mice (commercially availablefrom Taconic Farms, Inc, Hudson, N.Y. USA). Tumors were allowed to growfor 6 days. Mice were then distributed into groups of 10 mice based onranking of initial tumor volume to achieve groups in which the averagetumor size was 250 mm³ Example 5 was formulated in 2% HPMC(Hypermellose; HY122-13; commercially available from Spectrum ChemicalManufacturing Gardena, Calif. USA)/1% Tween-80 (Crillet 4 HP;commercially available from Croda, Inc Edison, N.J. USA) and dosed dailyor twice daily 6 hours apart for a total of 10 days with 6.25 mg/kg,12.5 mg/kg, 25 mg/kg, 37.5 mg/kg, 50 mg/kg, 75 mg/kg or 150 mg/kg.Tumors were measured every other day using two-dimensional calipers andtumor volumes were estimated using the equation Width²×Length×0.5. FIG.1 shows the calculated tumor volume as a function of time after twicedaily dosing with Example 5. Dose-dependent inhibition of tumor growthwas observed. All doses were statistically different from the vehicletreated group. Statistical significance was evaluated using RMANOVA onlog transformed tumor volume with baseline as covariate. P values areshown in FIG. 1.

Tumor Growth Inhibition in Colo205 Xenograft Tumors

The activity of Example 5 was evaluated in the human colon carcinoma,Colo205, xenograft tumor model. This cell line expresses CDK4/6 but notFLT3. Therefore, this system will measure the activity of Example 5against CDK4/6. Colo205 cells (obtained from American Type CultureCollection, Manassas, Va., USA) were cultured in DMEM cell growth mediumcontaining 10% fetal bovine serum (commercially obtained fromInvitrogen, Carlsbad, Calif. USA). Two million cells were innoculated onthe right flank of CrTac:NCR-Foxn1^(nu) nude mice (commerciallyavailable from Taconic Farms, Inc, Hudson, N.Y. USA) and allowed to growfor 13 days. Mice were then distributed into groups of 10 mice based onranking of initial tumor volume to achieve groups in which the averagetumor size was 170 mm³ Example 5 was formulated in 2% HPMC(Hypermellose; HY122-13; commercially available from Spectrum ChemicalManufacturing Gardena, Calif. USA)/1% Tween-80 (Crillet 4 HP;commercially available from Croda, Inc Edison, N.J. USA) and dosed dailyor twice daily 6 hours apart for a total of 10 days with 12.5 mg/kg, 25mg/kg, 37.5 mg/kg, or 50 mg/kg. Tumors were measured every other dayusing two-dimensional calipers and tumor volumes were estimated usingthe equation Width²×Length×0.5. FIG. 2 shows the calculated tumor volumeas a function of time after twice daily dosing with Example 5.Dose-dependent inhibition of tumor growth was observed. All doses werestatistically different from the vehicle treated group. Statisticalsignificance was evaluated using RMANOVA on log transformed tumor volumewith baseline as covariate. P values are shown in FIG. 2.

All publications and patent applications cited in this specification arehereby incorporated by reference herein in their entireties and for allpurposes as if each individual publication or patent application werespecifically and individually indicated as being incorporated byreference and as if each reference was fully set forth in its entirety.Although the foregoing invention has been described in some detail byway of illustration and example for purposes of clarity ofunderstanding, it will be readily apparent to those of ordinary skill inthe art in light of the teachings of this invention that certain changesand modifications may be made thereto without departing from the spiritor scope of the appended claims.

1. A compound of Formula I:

or a pharmaceutically acceptable salt thereof, a hydrate thereof, or amixture thereof, wherein: R¹ is a group of Formula IA, Formula IB,Formula IC, or Formula ID

wherein the

symbol indicates the point of attachment of the group of Formula IA, IB,IC, or ID to the rest of the molecule; R² is a C₅-C₇ cycloalkyl group,is a 5 to 7-membered heterocyclyl group that includes 1, 2, or 3heteroatoms selected from N, O, and S, or is a C₇-C₁₀ bicyclic group;wherein the C₅-C₇ cycloalkyl group, the 5 to 7 membered heterocyclylgroup, or the C₇-C₁₀ bicyclic group is unsubstituted or is substitutedwith 1-3 substituents independently selected from unsubstituted —(C₁-C₆alkyl), —OH, halo, —O—(C₁-C₆ alkyl), —CO₂H, —C(═O)—O—(C₁-C₆ alkyl),—C(═O)—NR′R″, —NR′R″, or a substituted —(C₁-C₄ alkyl), wherein thesubstituted —(C₁-C₄ alkyl) is substituted with 1-3 substituentsindependently selected from halo, —OH, —OCH₃, —S(═O)₂—CH₃, or—C(═O)—CH₃; R^(3a) is selected from —H, —F, or —Cl, —(C₁-C₃ alkyl), or—O—(C₁-C₃ alkyl); R^(3b) is —H, halo, —OH, —O—(C₁-C₆ alkyl),unsubstituted —(C₁-C₆ alkyl), —NR′R″, —C(═O)—(C₁-C₆ alkyl),—C(═O)—O—(C₁-C₆ alkyl), —C(═O)—NR′R″, or a substituted —(C₁-C₆ alkyl),wherein the substituted —(C₁-C₆ alkyl) is substituted with 1-3substituents independently selected from halo, —OH, —OCH₃, —CN, or —NO₂;R^(3c) is —H, —(C₁-C₃ alkyl), or halo; R⁴ is —H; R⁵ is —H; R⁶ isselected from —H, —(C₁-C₆ alkyl), —C(═O)—(C₁-C₆ alkyl), —C(═O)—O—(C₁-C₆alkyl), —C(═O)—C(═O)—OH, —C(═O)—NR′R″, or —S(═O)—NR′R″, wherein thealkyl group of the —(C₁-C₆ alkyl), —C(═O)—(C₁-C₆ alkyl), and—C(═O)—O—(C₁-C₆ alkyl) groups is unsubstituted or is substituted with1-3 substituents independently selected from —OH, F, —S(═O)₂—(C₁-C₆alkyl), —O—(C₁-C₆ alkyl), —NR′R″, or —CN; R^(7a) is —H, —CH₃, or halo;R^(7b) is —H, —(C₁-C₆ alkyl), or halo; or R^(7b) is absent if R¹ is agroup of Formula IB or Formula ID; R^(7c) is —H, unsubstituted —(C₁-C₆alkyl), halo, —O—(C₁-C₆ alkyl), —NO₂, —CN, —NR′R″, —CO₂H,—C(═O)—O—(C₁-C₆ alkyl), —C(═O)—NR′R″, or a substituted —(C₁-C₆ alkyl),wherein the substituted —(C₁-C₆ alkyl) is substituted with 1-3substituents independently selected from —OH, halo, —O—(C₁-C₆ alkyl),—CN, —NR′R″, or —S(═O)₂—CH₃; or R^(7c) is absent if R¹ is a group ofFormula IA or Formula IC; R^(8a) is —H, unsubstituted —(C₁-C₆ alkyl), ora substituted —(C₁-C₆ alkyl), wherein the substituted —(C₁-C₆ alkyl) issubstituted with 1-3 substituents independently selected from —OH, halo,or —O—(C₁-C₆ alkyl); R^(8b) is —H, unsubstituted —(C₁-C₆ alkyl), or asubstituted —(C₁-C₆ alkyl), wherein the substituted —(C₁-C₆ alkyl) issubstituted with 1-3 substituents independently selected from —OH, halo,or —O—(C₁-C₆ alkyl); or R^(8a) and R^(8b), when taken together, canrepresent ═O; R^(8c) is selected from —H, —OH, unsubstituted —(C₁-C₆alkyl), or a substituted —(C₁-C₆ alkyl), wherein the substituted —(C₁-C₆alkyl) is substituted with 1-3 substituents independently selected from—OH, halo, or —O—(C₁-C₆ alkyl); R^(8d) is —H, unsubstituted —(C₁-C₆alkyl), or a substituted —(C₁-C₆ alkyl), wherein the substituted —(C₁-C₆alkyl) is substituted with 1-3 substituents independently selected from—OH, halo, or —O—(C₁-C₆ alkyl); R^(8e) is —H, unsubstituted —(C₁-C₆alkyl), or a substituted —(C₁-C₆ alkyl), wherein the substituted —(C₁-C₆alkyl) is substituted with 1-3 substituents independently selected from—OH, halo, or —O—(C₁-C₆ alkyl); R^(8f) is —H, unsubstituted —(C₁-C₆alkyl), or a substituted —(C₁-C₆ alkyl), wherein the substituted —(C₁-C₆alkyl) is substituted with 1-3 substituents independently selected from—OH, halo, or —O—(C₁-C₆ alkyl); or R^(8e) and R^(8f), when takentogether, can represent ═O; and R′ and R″ are independently selectedfrom —H, unsubstituted —(C₁-C₄ alkyl), or —(C₁-C₄ alkyl) substitutedwith 1 to 3 substituents independently selected from —OH or —F.
 2. Thecompound of claim 1 or the pharmaceutically acceptable salt thereof, thehydrate thereof, or the mixture thereof, wherein R² is a C₅-C₇cycloalkyl group that is unsubstituted or is substituted with 1-3substituents independently selected from unsubstituted —(C₁-C₆ alkyl),—OH, halo, —O—(C₁-C₆ alkyl), —CO₂H, —C(═O)—O—(C₁-C₆ alkyl),—C(═O)—NR′R″, —NR′R″, or a substituted —(C₁-C₄ alkyl), wherein thesubstituted —(C₁-C₄ alkyl) is substituted with 1-3 substituentsindependently selected from halo, —OH, —OCH₃, —S(═O)₂—CH₃, or—C(═O)—CH₃.
 3. The compound of claim 2 or the pharmaceuticallyacceptable salt thereof, the hydrate thereof, or the mixture thereof,wherein R² is a cyclohexyl group substituted with a —(C₁-C₂ alkyl)group.
 4. The compound of claim 3 or the pharmaceutically acceptablesalt thereof, the hydrate thereof, or the mixture thereof, wherein R² isa group of formula

wherein the

symbol indicates the point of attachment to the rest of the molecule. 5.The compound of claim 1 or the pharmaceutically acceptable salt thereof,the hydrate thereof, or the mixture thereof, wherein R¹ is a group ofFormula IA or IB.
 6. The compound of claim 5 or the pharmaceuticallyacceptable salt thereof, the hydrate thereof, or the mixture thereof,wherein R¹ is a group of Formula IA.
 7. The compound of claim 1,wherein: R² is a C₅-C₇ cycloalkyl group that is unsubstituted or issubstituted with 1-3 —(C₁-C₆ alkyl) groups; R^(3a) is selected from —H,—(C₁-C₃ alkyl), or —O—(C₁-C₃ alkyl); R^(3b) is —H; R^(3c) is —H; R⁴ is—H; R⁵ is —H; R⁶ is selected from —H, —(C₁-C₆ alkyl), —C(═O)—(C₁-C₆alkyl), or —C(═O)—C(═O)—OH, wherein the alkyl group of the —(C₁-C₆alkyl) and —C(═O)—(C₁-C₆ alkyl) groups is unsubstituted or issubstituted with 1-3 substituents independently selected from —OH, F,—S(═O)₂—(C₁-C₆ alkyl), or —O—(C₁-C₆ alkyl); R^(7a) is —H; R^(7b) is —H;or is absent if R¹ is a group of Formula IB or Formula ID; R^(7c) is —H;or is absent if R¹ is a group of Formula IA or Formula IC; R^(8a) is —H;R^(8b) is —H; R^(8c) is selected from —H, —OH, or unsubstituted —(C₁-C₆alkyl); R^(8d) is —H; R^(8e) is —H; and R^(8f) is —H, or thepharmaceutically acceptable salt thereof, the hydrate thereof, or themixture thereof.
 8. The compound of claim 1, wherein the compound hasthe Formula IIA

or the pharmaceutically acceptable salt thereof, the hydrate thereof, orthe mixture thereof, wherein: R^(3a) is selected from —H, —F, or —Cl,—(C₁-C₃ alkyl), or —O—(C₁-C₃ alkyl); R^(3b) is —H, halo, —OH, —O—(C₁-C₆alkyl), unsubstituted —(C₁-C₆ alkyl), —NR′R″, —C(═O)—(C₁-C₆ alkyl),—C(═O)—O—(C₁-C₆ alkyl), —C(═O)—NR′R″, or a substituted —(C₁-C₆ alkyl),wherein the substituted —(C₁-C₆ alkyl) is substituted with 1-3substituents independently selected from halo, —OH, —OCH₃, —CN, or —NO₂;R⁶ is selected from —H, —(C₁-C₆ alkyl), —C(═O)—(C₁-C₆ alkyl),—C(═O)—C(═O)—OH, —C(═O)—NR′R″, or —S(═O)—NR′R″, wherein the alkyl groupof the —(C₁-C₆ alkyl) and —C(═O)—(C₁-C₆ alkyl) groups is unsubstitutedor is substituted with 1-3 substituents independently selected from —OH,F, —S(═O)₂—(C₁-C₆ alkyl), —O—(C₁-C₆ alkyl), —NR′R″, or —CN; and R^(8c)is selected from —H, —OH, unsubstituted —(C₁-C₆ alkyl), or a substituted—(C₁-C₆ alkyl), wherein the substituted —(C₁-C₆ alkyl) is substitutedwith 1-3 substituents independently selected from —OH, halo, or—O—(C₁-C₆ alkyl).
 9. The compound of claim 8, wherein: R^(3a) isselected from —H, —(C₁-C₃ alkyl), or —O—(C₁-C₃ alkyl); R^(3b) is —H; R⁶is selected from —H, —(C₁-C₆ alkyl), —C(═O)—(C₁-C₆ alkyl), or—C(═O)—C(═O)—OH, wherein the alkyl group of the —(C₁-C₆ alkyl) and—C(═O)—(C₁-C₆ alkyl) groups is unsubstituted or is substituted with 1-3substituents independently selected from —OH, F, —S(═O)₂—(C₁-C₆ alkyl),or —O—(C₁-C₆ alkyl); and R^(8c) is selected from —H, unsubstituted—(C₁-C₆ alkyl), or —OH, or the pharmaceutically acceptable salt thereof,the hydrate thereof, or the mixture thereof.
 10. The compound of claim 9or the pharmaceutically acceptable salt thereof, the hydrate thereof, orthe mixture thereof, wherein R^(8c) is selected from —H, —CH₃, or —OH.11. The compound of claim 10 or the pharmaceutically acceptable saltthereof, the hydrate thereof, or the mixture thereof, wherein R^(8c) is—H.
 12. The compound of claim 11 or the pharmaceutically acceptable saltthereof, the hydrate thereof, or the mixture thereof, wherein R^(3a) is—H.
 13. The compound of claim 12 or the pharmaceutically acceptable saltthereof, the hydrate thereof, or the mixture thereof, wherein R⁶ isselected from —H, —C(═O)—CH₃, —CH₂CH₂OH, —CH₂CH₂CH₂OH, —C(═O)—CH₂OH,—C(═O)—C(═O)—OH, —CH₂CH₂CF₃, —CH₂CH₂F, —CH₂CH₂S(═O)₂—CH₃, or—CH₂CH₂OCH₃.
 14. The compound of claim 13 or the pharmaceuticallyacceptable salt thereof, the hydrate thereof, or the mixture thereof,wherein R⁶ is selected from —C(═O)—CH₃ or —C(═O)—CH₂OH.
 15. A compoundselected from

or a pharmaceutically acceptable salt or a hydrate thereof.
 16. Thecompound of claim 15, wherein the compound is

or the pharmaceutically acceptable salt or the hydrate thereof.
 17. Thecompound of claim 15, wherein the compound is

or the pharmaceutically acceptable salt or the hydrate thereof.
 18. Thecompound of claim 15, wherein the compound is

or the pharmaceutically acceptable salt or the hydrate thereof.
 19. Acompound selected from

or a pharmaceutically acceptable salt or a hydrate thereof.
 20. Thecompound of claim 1 in a neutral form.
 21. The pharmaceuticallyacceptable salt or the hydrate thereof of claim
 1. 22. Thepharmaceutically acceptable salt or the hydrate thereof of claim 21,wherein the pharmaceutically acceptable salt or the hydrate thereof isselected from a chloride salt, a methanesulfonate salt, or abenzenesulfonate salt.
 23. A pharmaceutical composition, thepharmaceutical composition comprising a therapeutically effective amountof the compound, the pharmaceutically acceptable salt, the hydratethereof, or the mixture thereof according to claim 1 and at least onepharmaceutically acceptable excipient, carrier, or diluent.
 24. Thecompound of claim 15 in a neutral form.
 25. The pharmaceuticallyacceptable salt or the hydrate thereof of claim
 15. 26. Thepharmaceutically acceptable salt or the hydrate thereof of claim 25,wherein the pharmaceutically acceptable salt or the hydrate thereof isselected from a chloride salt, a methanesulfonate salt, or abenzenesulfonate salt.
 27. A pharmaceutical composition, thepharmaceutical composition comprising a therapeutically effective amountof the compound, the pharmaceutically acceptable salt, or the hydratethereof according to claim 15 and at least one pharmaceuticallyacceptable excipient, carrier, or diluent.
 28. The compound of claim 19in a neutral form.
 29. The pharmaceutically acceptable salt or thehydrate thereof of claim
 19. 30. The pharmaceutically acceptable salt orthe hydrate thereof of claim 29, wherein the pharmaceutically acceptablesalt or the hydrate thereof is selected from a chloride salt, amethanesulfonate salt, or a benzenesulfonate salt.
 31. A pharmaceuticalcomposition, the pharmaceutical composition comprising a therapeuticallyeffective amount of the compound, the pharmaceutically acceptable salt,or the hydrate thereof according to claim 19 and at least onepharmaceutically acceptable excipient, carrier, or diluent.
 32. A methodof treating cancer, the method comprising: administering to a subject aneffective amount of the compound, the pharmaceutically acceptable saltthereof, the hydrate thereof, or the mixture thereof of claim
 1. 33. Themethod of claim 32, wherein the cancer is selected from acute myeloidleukemia, acute lymphoblastic leukemia, myelodysplastic syndrome,multiple myeloma, chronic myeloid leukemia, acute lymphocytic leukemia,chronic lymphocytic leukemia, non-Hodgkin lymphoma, another lymphoma,another myeloma, or another leukemia.
 33. The method of claim 33,wherein the cancer is acute myeloid leukemia.
 34. The method of claim32, wherein the cancer is selected from breast cancer, colorectalcancer, small cell lung carcinoma, head and neck, glioblastoma,pancreatic, gastrointestinal, liver, prostate, ovarian, testicular,endometrial, bladder, melanoma, osteosarcoma, or another sarcoma. 35.The method of claim 32, wherein the cancer is Rb-positive.
 36. A methodof treating cancer, the method comprising: administering to a subject(a) an effective amount of the compound, the pharmaceutically acceptablesalt thereof, the hydrate thereof, or the mixture thereof of claim 1;and (b) at least one second therapeutic agent used in the treatment ofcancer.
 37. The method of claim 36, wherein the second therapeutic agentis cytosine arabinoside, daunorubicin, idarubicin, doxorubicin,cyclophosphamide, etoposide, carboplatin, fludarabine, mitoxantrone,dexamethasone, rituximab, midostaurin, a granulocyte colony-stimulatingfactor, filgrastim, PEG-filgrastim, lenograstim, decitabine,azacitidine, paclitaxel, gemcitibine, motesanib disphosphate,panitumumab, an antibody directed against CD33, or a CD33 bispecificT-cell engager antibody.